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Echo 500 Series User Manual Software version 2.3

1. 6 7 6 2 3 Clean and refill the coupling fluid bottle 6 9 6 2 4 Empty the coupling fluid bottle 6 10 6 2 5 Replace the water filter 6 14 6 2 6 Clean the anti static bars 6 15 6 2 7 Replace the AC power fuse 6 17 6 3 Extended Non Use and Storage 6 18 6 3 1 Maintenance During Extended Non Use 6 18 6 3 2 Preparation for storage 6 19 Chapter 7 Contact Information and Troubleshooting 7 14 Contact information SEUL sk BS ANS de ERA Ge ete Ta 4 7 1 7 2 Setup problems Lane aa na ee dey at e AR Because 7 2 7 3 Maintenance alerts 3445 pe sion ee No a ares 4 7 3 Appendix A ADE Technology Aa ADE RIStON dnan RER RES AT esse Sewn A 1 A2 ADE and the Echo liquid handler A 2 A3 Source microplate survey A 3 AA Fluid transteran atures sate le MA EDS EVENE De A 6 A 4 1 Positioning the ADE elements A 6 A 4 2 Creating the acoustic pulse A 6 A 4 3 Transferring the droplet A 7 A 4 4 Examples of acoustic droplet ejection A 8 Appendix B Barcode Locations B 1 Short flange height microplates B 2 B 2 Medium flange height microplates B 2 Appendix C Chiller Information CAL Shipping CONten
2. 2 11 2 4 Instrument Relocation 2 12 2 4 1 Prepare the Echo liquid handler foramove 2 12 2 4 2 Reinstall the Echo liquid handler after a move 2 13 Chapter 3 System Description 3 1 System overview se ie oe died we ane we wee oe Pw ws 3 2 3 1 1 Echo front panel Lana mama Ra lek che hs tiens 3 2 3 1 2 Echo liquid handler back panel 3 4 3 2 Component description 29 ge Le NE beled te Lite 3 5 3 21 Process DOME cet ccd hye Ses tytn hands ray aie 3 5 3 2 2 Source plate gripper stage 3 6 3 2 3 Source plate insert 2 54 Ose aoe haie Ris 3 6 3 2 4 Destination plate gripper stage 3 8 3 2 5 Status indicator lights 3 9 3 20 ECD Screen See le RER RAA TR ee TA SRE 3 10 22h EMO SWitC h eloi Ser rete ea aa ea erg eres car et 3 11 3 2 8 Anti static bars cnc 0 0 asdcxw aoben 268 v8 eek wae ea 3 11 3 2 9 AC power and fuse compartment 3 13 3 2 10 Coupling fluid bottle 25848 sv ed eo eS 3 15 3 2 11 Fluid chiller eee EN EN ee Rae Re Bee 3 16 S212 SUIGEAANK 51m oo aie a eS weasel a wo mee wees 3 16 3 2 13 Fluidics panel EN Rs AlN arene eR 3 17 3 3 Specifications Sep An da sn ds An A sn nd 3 18 33 PAVSI A RE es Neue et Re EN NS teat gh A 3 18 3 3 2 Mechanical DER yf See dp ENS Te dons 3 18 3 3 3 Environmental sie we eae il ado nee eue 3 19 3 3 4 Eleal en a te
3. Q The waste water bottle is full on older systems only See Waste bottle indicator on page 5 43 Q The process door is neither fully open nor fully closed Check for obstruction Check compressed air pressure it could be too low See Compressed air pressure indicator on page 5 43 Q The upper anti static bar is neither fully up nor fully down Q The source plate dryer nozzle is neither fully up nor fully down Q Fault light is red Possible causes Q The user supplied compressed air supply is low lt 70 PSI or nonexistent Check compressed air pressure See Compressed air pressure indicator on page 5 43 Check compressed air pressure See Compressed air pressure indicator on page 5 43 See Status indicator lights on page 3 8 Increase air pressure See Compressed air pressure indicator on page 5 43 Contact Information and Troubleshooting 1 3 Table 7 1 Echo Maintenance Alerts Maintenance Alert Procedure Q The user supplied vacuum pressure is low lt 15 in Hg Vac or nonexistent Q The EMO Emergency Motion Off Increase vacuum pressure See Vacuum supply pressure indicator on page 5 43 See EMO indicator on page 5 42 switch has been activated Q Acoupling fluid leak has been detected Q LCD message indicates user assistance needed Q Client PC indicates a problem Se
4. Chapter 6 Maintenance and Service covers maintenance schedules and user level preventive maintenance procedures Chapter 7 Contact Information and Troubleshooting provides setup problems and signal alerts Appendix A ADE Technology describes acoustic droplet ejection technology Appendix B Barcode Locations specifies where a microplate barcode label must be placed to be read by the Echo liquid handler Appendix C Chiller Information contains chiller descripton setup and operation and troubleshooting Echo Liquid Handler User Manual Conventions used in this manual Some of the text in this manual uses special formatting to help indicate emphasis or keystrokes The text conventions are as follows Convention Small caps bold Courier Blue underlined Quotation marks Example Press the NEXT button client txt client old2 txt Software installation See Manage labware definitions on page 5 31 Meaning Indicates an on screen button label menu title or menu item Indicates a display output printed output keyboard input or file names Signifies a hyperlink to a topic Refers you to a topic elsewhere in the manual Usually includes a hyperlink Preface xix XX Echo Liquid Handler User Manual CHAPTER INTRODUCTION The Labcyte Echo 500 series includes the Echo 520 550 and 555 liquid handlers Throughout this manual the des
5. gt 3 14004 New Focus ToF us 33 377 33 593 Previous New us 0 2236 0 00765 Amplitude v 2 971 280195 2 63291 31 2797 32 2604 33 241 34 2217 35 2023 36 183 ToF us Press Finish to save the new Focus ToF Press Back to retract source gripper Focal Sweep Focal sweep is a term used to describe sweeping or moving the acoustic transducer in a predefined Z vertical range while capturing a series of acoustic echoes from the plate bottom and potentially the well fluid surface if the well is not empty LS Note The focal sweep tool is used for diagnostics purposes only Use this tool only when directed by Labcyte personnel 1 Create a calibration plate each well containing 20 50 ul of fluid 70 100 DMSO water 2 In the CALIBRATION window click the FOCAL SWEEP button 3 Set up the FOCAL SWEEP PARAMETERS Wells Select which wells will be included in the focal sweep Deselect All Deselects all previously selected wells Start Stop ToF Defines the start and stop points of the focal sweep in terms of BB ToF well plate bottom time of flight in microseconds Labcyte recommends a START TOF of 32 5 us and a STOP TOF of 34 5 us Plate Name Select the type of source plate being used Its definition and name must already be stored in the Echo database Z Increment Define the acoustic transducer s Z motion resolution in microns Labcyte recommends a Z INCREMENT of 50 um Echo Liquid Handler S
6. m If the Echo liquid handler will be disconnected and stored for an indefinite period of time then the instrument must be prepared for storage See section 6 3 2 6 3 1 Maintenance During Extended Non Use The following procedure assumes that the Echo liquid handler is still connected to the power source chiller air and vacuum lines 1 Recirculate the coupling fluid once a week See the procedure Cycle fluid through the Echo system on page 6 12 2 Perform scheduled maintenance at two week and one month intervals Call Labcyte Service and Support to schedule six month maintenance See Maintenance schedule on page 6 2 6 18 Echo Liquid Handler User Manual 6 3 2 Preparation for storage Contact Labcyte Service and Support to arrange for the storage of the Echo liquid handler See Contact information on page 7 1 Include an estimate of how long the instrument will be stored and where it will be stored The Labcyte service engineer may replace the coupling fluid with fresh water and algaecide and seal the nose cone to keep the acoustic transducer wet during storage When you are ready to return the Echo to full operation contact Labcyte Service and Support The Labcyte service engineer will determine what assembly or testing steps are required to ensure that the instrument is operating correctly Note if you plan to store the Echo liquid handler in a different facility follow the moving procedur
7. m The Echo liquid handler has not completed its initialization e Check the gripper stage homing sequences particularly the theta rotation of the destination gripper stage e Check the door opening sequence m The Ethernet cable is not connected between the client PC and the Echo liquid handler m The Echo software is not configured to connect to the instrument IP address or hostname is missing or incorrect Check for the cause attempt to resolve the problem and try to launch the software again If the message persists call Labcyte Service and Support Echo initialization The Echo liquid handler performs a system initialization that includes the following actions m All three stages move to their home position e Source stage moves front to rear on its Y axis e Destination stage moves left to right on its X axis front to rear on its Y axis up and down on its Z axis and partially rotates on its Theta axis e Acoustic transducer stage moves left to right on its X axis front to rear on its Y axis and up and down on its Z axis 4 2 Echo Liquid Handler User Manual L Note The LCD screen reports each step of the homing sequence as it occurs Homing synchronizes the mechanical stages with the controlling software Process door opens Destination stage extends then retracts Process door closes Upper anti static bar moves to its uppermost position then both anti static bars turn off Plate dryer nozzle mo
8. Figure 5 2 Starting the Echo liquid handler from the Start menu E Adobe gt fin Echo 5XX b A Echo Liquid Handler Software E Microsoft Keyboard gt Echo Cheny Pick gt Release Notes fan Microsoft Office Tools gt The Echo software connects to the Echo liquid handler and displays the following screens Figure 5 3 Echo liquid handler welcome screen LABCYTE Echo Liquid Handler Software Echo 520 Liquid Handler Echo 550 Liquid Handler Echo 555 Liquid Handler copyright 2005 2008 This program is protected by U S and intemational copyright laws as described in the About Box 5 2 Echo Liquid Handler User Manual Figure 5 4 Echo liquid handler protocol screen LABCYTE gt Protocol List 384 10nL 5 copies 10 0nl 384 384 384PP Aqueous buf F protein to 1536LDY 5 copies 2 5nl 384 1536 I 384PP DMSO to 384P5 10 copies 5 0nl 384 384 O Note If the Echo software cannot connect with the Echo liquid handler the following message will appear Figure 5 1 Connection failure message Failed To Connect To Echo Server i xj i Host refused connection Verify address and port number localhost Click OK to try to connect to server again click Cancel to exit Echo Liquid Handler This message may appear when the Echo software attempts to connect to the Echo liquid handler during system startup Possible causes for connection failure include m The Echo liquid handler has not been turne
9. LABCYTE Liquid Handler 500 Series User Manual Software version 2 3 Echo Liquid Handler User Manual Labcyte Part Number 001 5331 Rev4 January 2011 Copyright 2011 Labcyte Inc All rights reserved The information contained in this manual is subject to change without notice Any unauthorized changes or modifications to the Echo liquid handler will void its warranty Contact Labcyte Inc prior to making any change or modification Microsoft Windows Windows XP Activex Excel and Microsoft SQL Server are either registered trademarks or trademarks of Microsoft Corporation in the United States and other countries Intel Pentium Ill and Pentium 4 are trademarks or registered trademarks of Intel Corporation or its subsidiaries in the United States and other countries Alconox is a registered trademark of Alconox Inc Colder Products is a registered trademark of Colder Products Company All other registered or unregistered trademarks used herein are the exclusive property of their respective holders For research purposes only not for use in diagnostics The Echo liquid handler is covered by one or more of the following patents 6 416 164 6 548 308 6 603 118 6 612 686 6 642 061 6 666 541 6 707 038 6 710 335 6 746 104 6 802 593 6 808 934 6 809 315 6 849 423 6 855 925 6 869 551 6 893 836 6 893 115 6 916 083 6 932 097 6 938 987 6 938 995 6 991 917 7 070 260 7 090 333 7 354 141 7 405
10. Q Check the coupling fluid bottle level daily e f the coupling fluid level is at 350 mL or less clean and refill the coupling fluid bottle and add algaecide See Refill the coupling fluid bottle on page 6 7 e f the coupling fluid is cloudy clean and refill the coupling fluid bottle and add algaecide See Clean and refill the coupling fluid bottle on page 6 9 e Check the waste bottle daily older Echo systems only Empty it if it is greater than half full See Empty and clean the waste water bottle on page 6 13 Q Check the plate insert for proper positioning or damage See Source plate insert on page 3 6 6 2 Echo Liquid Handler User Manual 6 1 2 Echo maintenance alerts The Echo liquid handler contains internal checks to ensure that it performs liquid fills under optimal conditions If for example the coupling fluid temperature is outside the normal range 21 2 C 23 0 C a yellow warning light appears on the Echo front and back panels If the coupling fluid temperature continues to rise or fall outside of the warning range 20 2 C 24 0 C a red fault light appears When you see the warning or fault lights click the Diagnostics tab in the Echo software to view the specific alert Figure 6 1 Diagnostic window Protocols Labware Diagnostics Calibration Advanced Device control and status Door J Open O Source
11. on page 6 10 and Cycle fluid through the Echo system on page 6 12 Maintenance and Service 6 7 4 Visually inspect the used coupling fluid It should not be cloudy or visually contaminated with algae or other substances If it is clean the bottle see Clean and refill the coupling fluid bottle on page 6 9 before refilling it 5 Fill the coupling fluid bottle with 900 mL of distilled water and add 1 drop 50 uL of algaecide CS Note The ratio of algaecide to water is optimized for deterring the growth of algae while minimizing residual moisture on the bottom of the plate 6 Replace the bottle cap Hand tighten the cap securely to ensure a tight seal 7 Carefully inspect the coupler fluid tube and attached flexible tubing for the presence of bubbles If you see bubbles you will need to purge then recycle the fluid through the system after you complete this procedure See Purge the fluid circuit on page 6 10 and Cycle fluid through the Echo system on page 6 12 8 Reconnect the vacuum line and turn the coupling fluid pump on through the DIAGNOSTIC window L Note The surge tank will add a few seconds to starting the vacuum A Caution Make sure that the vacuum line is connected and working correctly before turning the coupling fluid pump back on If the vacuum line is disconnected or the vacuum system is not operating correctly or the cap on the fluid bottle is not tightly sealed when the
12. 2 15 0 984 2 5 132 2 053 77 7 B4 1826 3 2 15 0 984 7 5 132 2 053 77 77 B5 1 826 3 12 15 0 984 77 2 5 132 2 053 2 9 3 3 B6 1 826 2 15 10 984 7 15 132 2 053 7 B 1 826 2 15 0 984 7 15 132 2 053 7 nasal an an gt ADNA an 9 To save the survey data to the client PC click the FILE button and select from the following choices Eluid Composition cs Fluid Thickness cs Volume csv m SAVE FLUID COMPOSITION CSV 1 m SAVE FLUID THICKNESS CSV B m SAVE RAW csv E Raw csw Note that FILE SAVE FLUID COMPOSITION will save raw processed homogeneous or processed inhomogeneous survey data depending on which POST PROCESSING radio button is selected The standard Windows SAVE AS dialog box is displayed Figure 5 2 Diagnostic survey file saved El File name 0 07 22 15 52 42 platesurvey fluid thickness Save as type CSV Files csv v Cancel 5 50 Echo Liquid Handler User Manual 5 8 Calibration window Several Echo mechanisms can be user calibrated The following motion power acoustic transducer and barcode scanner related calibration procedures are found in the CALIBRATION window Figure 5 3 Calibration window Protocols Labware Diagnostics Calibration Advanced Motion Home Register P
13. 25 4 cm 10 in top clearance and sufficient front clearance to load and unload microplates by hand or by robot See the illustration on page 2 2 Weight 128 kg 283 lbs 3 3 2 Mechanical Air pressure Clean dry air at 552 kPa 80 PSI minimum 1034 kPa 150 PSI maximum Air connection Male coupling insert LC Series 14 NPT male straight thru and coupling body LC series in line ferruleless fitting for 6 4 mm 1 in ID tubing included in Echo liquid handler accessory kit Air consumption lt 28 3 liter min 1034 kPa lt 1 cf m 150 PSI Vacuum source e House vacuum Minimum vacuum at Echo connection 200 Torr 266 mbar 22 in Hg Vac Vacuum system capacity steady state 15 L min 0 9 m h 0 53 cf m Drying surge 80 L min for 10 sec 4 8 m h 2 8 cf m 33 max duty cycle e Vacuum pump Minimum vacuum at Echo connection 100 Torr 133 mbar 26 in Hg Vac Pump speed with 10 L surge tank 40 L min 2 4 m h 1 4 cf m without surge tank 80 L min 4 8 m h 2 8 cf m L Note If the vacuum measured at the Echo connection dips below 1 2 an atmosphere 380 Torr 15 in Hg Vac then the Echo liquid handler could go into an error state To ensure consistent vacuum Labcyte recommends the use of a 10 L surge tank contact your Labcyte representative 3 18 Echo Liquid Handler User Manual Vacuum connection Male coupling insert HFC Series 3 8 NPT male straight thru and coupling body HFC12 series in l
14. following actions occur A portion of the acoustic energy is absorbed by the material it is passing through m The ping encounters a change in impedance between two materials and some of the energy is reflected back toward its source m The remaining portion is transmitted into the next material The reflected sound waves are captured by the acoustic transducer Three signals are detected BB Bottom of the Bottom The BB echo is created by the impedance difference between the distilled water coupling fluid and the microplate material polypropylene cyclic olefin copolymer and so on m TB Top of the Bottom The TB echo is created by the impedance difference between the microplate material and the fluid in the microplate well m SR Surface Reflection The SR echo is created by the impedance difference between the microplate well fluid and the ambient air above its meniscus A 3 Figure A 1 The BB TB and SR echoes TB SR Bottom of the Bottom Top of the Bottom Surface Reflection Echo signal returned Echo signal returned Echo signal returned from the interface of from the interface of from the interface of the coupling fluid and the top of the the fluid s meniscus the microplate s microplate bottom and the air above it bottom and the fluid in the microplate well In Figure A 2 the echo signals are clearly distinguished This figure is only an example actual timing depends on the fill height in the well flui
15. or every other dest well 1536 well 1536 well standard 1 src plate to 1 dest plate 1 Destacking transferring liquid from a single source src plate to several destination dest plates 2 Stacking transferring liquid from several source plates to a single destination plate 1536 well 384 well CS Note See What types of liquid transfer are supported on page 1 11 for definitions of plate mapping terms Examples of protocols that cannot be created with the standard Echo software included with the Echo liquid handler installation m Transfers other than picture perfect For example well A1 into well C12 m Variable drop volume transfers For example 5 nL from source well A1 to destination well A1 10 nL from source well A2 to destination well A2 and so on m Transfers to non standard destination plates Note Complex and non standard liquid transfer protocols can be created by using the Echo Cherry Pick and Echo Plate Reformat applications or by programming your own custom protocols with the Echo ActiveX programming interface refer to the Labcyte Echo Integration Manual 1 Compound from the source plate wells are transferred to the same relative well positions in the des tination plate For example transferring compound from the first two columns in the source plate to the first two columns in the destination plate Echo Liquid Handler Software 5 11 5 5 1 Create a protocol To execute a liqu
16. the survey interval can be safely increased For example every 5 plates will cause a new survey to be performed when the source plate and first destination plate are loaded and every fifth destination plate thereafter Increasing the survey interval may improve throughput Default 1 CS Note The survey interval can be set to 0 however with this setting the protocol will not survey the plate Assume Homogeneous Plate When a source plate is surveyed to measure the DMSO water ratio in each well the Echo liquid handler creates a DMSO concentration map of the entire plate This map is post processed using a moving average filter if the ASSUME HOMOGENEOUS PLATE checkbox is selected There is no overhead associated with this processing Typically better liquid transfers are obtained using post processing when the source microplate contents are homogeneous A homogeneous source microplate is one where the solution to be transferred is very similar well to well J Assume Homogeneous Plate DMSO If the USE PREDEFINED checkbox is selected the Assume Homogeneous Plate feature is unavailable and its checkbox is grayed out Default Assume homogeneous plate off Power Monitoring Selecting the POWER MONITORING Rie checkbox causes the Echo liquid handler to monitor the RF power used to eject each fluid droplet Labcyte recommends that power monitoring not be enabled by the user unless directed to do so by Labcyte personne
17. yet ready for use Warning indicator The WARNING Status indicator reports conditions that will not allow normal operation Gray OK The Echo liquid handler is ready for use Yellow One or more of the following is true m The Echo stages are not homed See Home calibration on page 5 52 m The coupling fluid bottle level is low See Coupling fluid indicator on page 5 43 m The coupling fluid temperature is outside of the normal range 22 1 C 0 9 C 21 2 C 23 0 C The process door is neither fully open nor fully closed The upper anti static bar is neither fully up nor fully down The source plate dryer nozzle is neither fully up nor fully down Focus calibration is required Echo Liquid Handler Software 5 41 Fault indicator The Fault status indicator reports events or situations that must be responded to by the operator Normal operation may be affected or will be shortly Gray OK The Echo liquid handler is ready for use Red One or more of the following is true The user supplied compressed air supply is lt 552 kPa lt 80 psi See Compressed air pressure indicator on page 5 43 m The user supplied vacuum pressure is lt 78 kPa lt 15 in Hg Vac See Vacuum supply pressure indicator on page 5 43 m The coupling fluid temperature is outside of the warning range 20 2 C 24 0 C m The waste bottle is full See Waste bottle indicator on page 5 43 m The EMO switch has
18. 3 Source 1 Transfer Wells areg View Define Source 2 Transfer Wells A5 P8 View Source 3 Transfer Wells faci View Define Source 4 Transfer Wells AP View Define il lt Back Finish Cancel Create a new 1536 384 protocol definition Create a new 1536 384 protocol definition using the destacking features of the ADD New PROTOCOL WIZARD Compound from a single 1536 well source plate can be transferred to four 384 well destination plates See 384 plate to 1536 plate liquid transfer on page 1 14 for an illustration of plate mapping to more than one destination plates L Note ifthe new liquid transfer protocol requires a new destination microplate definition create it before running the ADD NEw PROTOCOL WIZARD See Manage labware definitions on page 5 31 1 Click the ADD button to start the ADD NEw PROTOCOL WIZARD 2 Protocol Name Enter a name for the new protocol It must not already be in use a Source Plate Type Select a source plate type from the first drop down list CS Note A 1536 well source microplate must already exist in the Echo database Contact Labcyte to purchase a 1536 well source plate definition See Contact information on page 7 1 b Destination Plate Type Select a destination plate type from the second drop down list The destination plate definition must already exist in the Echo database Echo Liquid Handler Software 5 19 Figure 5 1 Add new protocol wizard page
19. 3 Chiller Setup and Operation C 3 1 Setup Your Labcyte service representative will set up the chiller during Echo system installation If you need to set up the chiller again for example after relocation of the Echo system use the following procedure The Echo system should already be set up with tubing assembly attached to the fluid panel see Echo liquid handler user manual 1 Place the chiller on the benchtop near the Echo unit or on the floor under the Echo unit 2 Connect the Echo tubing assembly to the fluid connections on the chiller labeled Coolant Return for liquid returning to the chiller and Coolant Supply for fluid exiting the chiller Figure C 2 Tubing assembly connections COOLANT COOLANT RETURN SUPPLY 7 J 3 Connect the power supply cable m circular 2 pin connector to the 12 V DC 8 A on the chiller m three prong electrical connector to the electrical outlet C 3 C 3 2 Operation The Labcyte chiller temperature is controlled through three membrane keys below the 8 character LCD screen UP DOWN ENTER 1 Press the on off switch located next to the temperature display to turn on the chiller on off O 2 Set the temperature to 22 C a Press the UP or Down buttons to change the temperature values b Press the ENTER button to set the temperature Figure C 3 Chiller temperature setting DOWN ENTER The chiller will maintain the set temperature until it is cha
20. 4 12 Echo Liquid Handler User Manual 4 4 2 Start liquid transfer run You are ready to run the liquid transfer protocol 1 From the PROTOCOLS window select the protocol to be run If you move the mouse pointer over a protocol name a tool tip displays some of the protocol settings Figure 4 9 Protocols window LABCYTE gt l Protocol List EE ERE 384PP Aqueous buffiprotein to 15361 ri anie 384PP DMSO to 384P5 10 copieg Protocol Name 384 10nL Source Plate 334PP_4Q_SP Destination Plate 384PP _Dest Number of Copies 5 Transfer Volume 10 0nl Add Edit Remove 2 Click the RUN button You will be instructed to click the NEXT button The process door will open and the source plate gripper stage will extend outside of the Echo liquid handler Figure 4 10 Starting transfer protocol Run Protocol 384 10nL 5 copies 10 0nl 384 384 Press Next to start the transfer The door will open and the source gripper will be presented Manual Operation 4 13 3 Load the source microplate with well A1 in the inner left corner Click the Next button Figure 4 11 Loading source plate Run Protocol 384 10nL 5 copies 10 0nl 384 384 Please load source plate aggregate 1 of 1 Pressing Next will retract the source plate and present the destination gripper The source plate gripper stage retracts into the Echo liquid handler and the destination plate gripper stage is extended o
21. 6 3 The following table lists the normal maintenance alerts that a user would see as well as the respective corrective procedures To see a list of other factors that can trigger a Warning or Fault light see Chapter 7 Contact Information and Troubleshooting Table 6 2 Echo Maintenance Alerts fluid bottle Maintenance Alert Cause Procedure Q Warning light is yellow e The fluid level is low in the coupling Add more fluid See Refill the coupling fluid bottle on page 6 7 21 2 C 23 0 C e The coupling fluid temperature is outside of the normal range Check the chiller setting it should be at 22 C See Coupling fluid temperature and indicator on page 5 44 systems only e The waste bottle is full older Echo Q Fault light is red Empty the waste water bottle See Empty and clean the waste water bottle on page 6 13 nonexistent e The user supplied compressed air supply is low lt 70 PSI or Increase air pressure See Compressed air pressure indicator on page 5 43 20 2 C 24 0 C 6 4 Echo Liquid Handler User Manual e The coupling fluid temperature is outside of the warning range e The user supplied vacuum pressure is low lt 15 in Hg Vac or nonexistent Check the chiller setting it should be at 22 C See Coupling fluid temperature and indicator on page 5 44 Increase vacuum pressure See
22. Coupling fluid bottle Fluid chiller Fluidics panel m Specifications Physical Mechanical Environmental Electrical Communications Fluid transfer Client PC Supported labware Coupling fluid System Description 3 1 3 1 System overview This section provides the location and brief description of the components followed by detailed descriptions 3 1 1 Echo front panel Figure 3 1 Echo front panel Status indicators LCD screen Process door Emergency off switch shown open Upper anti static bar Destination plate gripper stage behind front cover Lower anti static bar Source plate gripper stage behind front cover with plate insert Barcode scanner optional behind front cover Process door The process door protects the operator from the moving mechanisms blocks stray light emitted by the barcode scanner lasers For more information see Process door on page 3 5 Destination plate gripper stage The destination plate gripper stage is the upper plate stage that extends from the Echo liquid handler The plate gripper end portion of the stage holds the destination microplate that will receive the liquid being transferred For more information see Destination plate gripper stage on page 3 7 Source plate gripper stage The source plate gripper stage is the lower plate stage that extends from the Echo liquid handler The plate gripper end portion of the stage h
23. Liquid Handler Software 5 39 Coupler pump on off The Echo liquid handler can be commanded to turn the coupler fluid pump on or off The associated indicator lights report whether the coupler fluid pump is turned on or off Coupler pump direction normal reverse The Echo liquid handler can be commanded to run its coupler fluid pump in the normal or reverse direction The associated indicator lights report the direction of coupler fluid pump flow L Note Do not run the pump in reverse for an extended period 5 40 Echo Liquid Handler User Manual 5 7 2 Status indicators The SYSTEM STATUS section of the DIAGNOSTICS window contains a vertical array of status indicators and readouts Four of them POWER WARNING FAULT and EMO correspond to the similarly named indicators on the Echo status panels Figure 5 1 System status section System Status Power Air Pressure 9 Warning VacuumPressure Fault Coupling Fluid EMO WasteBotte Fluid Leak Focus Calibration Coupling Fluid Temp 21 97 C RF Temp 1 43 45 C Power indicator The POWER status indicator provides more information than simply power on or power off Green The Echo liquid handler is powered on and is ready for use Gray One or more of the following conditions is true m The Echo liquid handler is not powered on m The Echo internal controller is not running m The Echo liquid handler is still initializing after being powered on and is not
24. Metric Allen wrenches 2 5 mm 3 mm and 4 mm Phillips screwdriver 2 Long needle nose pliers Part retrieval tool claw style retractable spring Lint free wipes Penlight flashlight Dental mirror Shop cloth to clean up spilled liquids Compressed air or vacuum with brush Materials required The following materials are required during Echo maintenance procedures m Spare Labcyte 1000 mL bottle square bottle catalog number ECHO AL15 S round bottle catalog number ECHO AL 15 R m Algaecide 4 mL bottle Labcyte catalog number ECHO AL04 A Caution When user level or more technical maintenance procedures are performed in a multiple client system take appropriate steps to avoid conflicts between clients such as turning on Maintenance Mode in the Diagnostic window 6 6 Echo Liquid Handler User Manual 6 2 2 Refill the coupling fluid bottle The Echo coupling fluid couples the energy from the acoustic transducer to the bottom of the source plate Bubbles in the tubing from very low fluid level or contamination in the fluid can reduce the accuracy or precision of source plate surveys and fluid transfers The coupling bottle should be refilled or cleaned and refilled immediately Maintaining clean coupling fluid at the proper fill level ensures a high level of performance in the Echo liquid handler Refill the coupling fluid bottle when one of the following occurs m COUPLING FLUID indicator on the Echo software DIAGNOSTICS t
25. Off Normally the Fault light is turned off indicating that the Echo liquid handler is functioning correctly m Red The Fault light is turned on when any of the following conditions are true e The user supplied compressed air supply is low lt 80 PSI or nonexistent See Compressed air pressure indicator on page 5 43 e The user supplied vacuum pressure is low lt 23 in Hg Vac or nonexistent See Vacuum supply pressure indicator on page 5 43 e The coupling fluid temperature is outside of the warning range 22 1 C 1 9 C 20 2 C 24 0 C e The waste water bottle is full on older Echo systems only See Waste bottle indicator on page 5 43 System Description 3 9 e The EMO Emergency Motion Off switch has been activated See EMO indicator on page 5 42 e A coupling fluid leak has been detected See Fluid leak indicator on page 5 42 3 2 6 LCD screen There is a four row LCD screen on the front of the Echo liquid handler and a duplicate on the rear panel They display messages that report what the Echo liquid handler is doing or what operator action is required Types of messages displayed include the following m Operating states m Procedure progress m Status m Requests for operator assistance 3 2 7 EMO switch The EMO Emergency Motion Off switch can be used in an emergency to shut down all mechanical Echo liquid handler activity There are two EMO switches one on the front panel a
26. POWER dialog box The Echo liquid handler performs the following procedure The most recent power calibration record stored in the Echo database is read This data will be used in the next step An RF pulse is generated Several RF measurements are taken A new amplifier feedback value is calculated and displayed as POWER AMPLIFIER FEEDBACK A new pulse energy value is measured and displayed as POWER PULSE ENERGY A new Vpp value is measured and displayed as POWER VPpP The new power calibration record is stored in the Echo database Figure 5 2 Calibrate Power dialog box after power calibration Calibrate Power Reference Amp 1 6 volts Pulse Energy 4 mJ r Power Amplifier Feedback 1 04069 Pulse Energy 2 77 mJ Vpp 86 1824 volts m System Gain 1 D Override Echo Liquid Handler Software 5 55 3 System Gain During normal operation the SYSTEM GAIN value should not be changed by the user However the user can override the automated power calibration by adjusting the SYSTEM GAIN scalar multiplier value The default SYSTEM GAIN value is 1 0 Select the OVERRIDE checkbox and enter a multiplier value manually 4 After power calibration has been performed and or the SYSTEM GAIN parameter has been modified there are three actions to choose from OK The OK button applies the new AMPLIFIER FEEDBACK value and or the new System Gain value and closes the CALIBRATE POWER
27. Vacuum supply pressure indicator on page 5 43 6 1 3 Scheduled maintenance Labcyte recommends that the following maintenance procedures be performed on a regular basis Maintenance promotes accurate precise and reliable fluid transfers Every two weeks Once every two weeks or earlier if the coupling fluid level becomes low the Echo user should perform the following preventive maintenance procedures m Purge the coupling fluid circuit clean the coupling fluid bottle and refill with algaecide water mixture A Caution Use the recommended amount of algaecide 50 uL algaecide 1000 mL distilled water Little or no algaecide results in algae growth which can decrease instrument performance and may require tubing replacement Excess algaecide can lead to fluid collection on the bottom of the microplate possible dripping and faster depletion of coupling fluid m Empty and clean the waste water bottle on older Echo systems only LS Note The waste water bottle should contain very little if any water If you are frequently emptying the waste water bottle contact Labcyte Service and Support Once per month Once per month the Echo user should perform the following procedure m Clean the anti static bar emitter points and ground reference surfaces with a soft bristled dry nylon toothbrush See Clean the anti static bars on page 6 15 m Change the water filter See Replace the water filter on page 6 14
28. a 100 120 VAC power source m 10A for a 200 240 VAC power source If you are unable to insert an AC plug into your AC receptacle contact an electrician to correct the situation Warning The Echo liquid handler operates with voltages and currents that can be lethal Pushing objects of any kind into the Echo liquid handler through slots or holes in its covers may cause serious electrical shock or may short out electrical circuits or parts Do not spill any liquid inside or on the Echo liquid handler Warning The Echo liquid handler contains user replaceable AC power fuses The fuse holder contains two fuses Use only the specified replacement fuses and the fuse replacement procedure found at Replace the AC power fuse on page 6 17 If a fuse requires repeated replacement the Echo liquid handler could have an electrical problem Do not use a malfunctioning Echo liquid handler Contact Labcyte Service and Support see Contact information on page 7 1 Warning Do not use AC power cords if the following conditions exist m The power cords are frayed or damaged m Other attached cords cables or receptacles are frayed or damaged Use of damaged power cords can cause an electrical shock hazard and result in severe injury Warning Do not connect plug in or disconnect unplug AC power cords if the following conditions exist m The Echo liquid handler or attached equipment has been exposed to excessive moisture or to liq
29. a protocol 5 12 5 5 2 Edit an existing protocol definition 5 24 5 5 3 Remove an existing protocol definition 5 25 5 5 4 Run a protocol 5 0 STE LM DRE RD eee ee 5 25 Labware window 4 4 LENS Gide qe ae draw LAN ae Goal SANS 5 28 Diagnostics WINDOW 15 inal tee eaten a waa al a ela 5 36 Dr DEVICE CONGO ES Re ER RARE NE oR Es von 5 37 5 7 2 Status indicators 420508 SS A Bh A DARA BS 5 41 5 7 3 Diagnostic procedures 5 46 Calibration window sae ia ve ti Mi AA s Re Ru 5 51 5 8 1 Motion calibration Ax one pone ne 5 52 5 8 2 Power calibration e aa VEN NUENE MERS 5 54 5 8 3 Transducer calibration 5 57 5 8 4 Barcode scanner calibration 5 61 Advanced window ea 44 4a eave NOX we SNES Here 5 63 5 9 1 Advanced liquid transfer controls 5 63 5 9 2 Advanced solvent concentration DMSO only 5 65 5 9 3 Advanced Echo control 44 44 444424444424 5 67 Contents V Chapter 6 Maintenance and Service 6 1 Maintenance schedule 0 nahn RH wR Re EAE Bee 6 2 6 1 1 Daily maintenance erreurs entered a a 6 2 6 1 2 Echo maintenance alerts 6 3 6 1 3 Scheduled maintenance 6 5 6 2 Maintenance procedures 6 6 6 2 1 Maintenance tools and materials 6 6 6 2 2 Refill the coupling fluid bottle
30. clicking the ABORT button 5 When the liquid transfer is complete click the NExT button Transfer for 1 of 1 complete Click Next to present destination gripper ESS ed a The destination plate gripper stage extends outside of the Echo liquid handler and reverts the plate to its upright position 6 Remove the destination microplate and click the Next button Run Protocol 384 10 nL 1 copy 10 0nl 384 384 Please remove destination plate 1 of 1 from the gripper The destination plate gripper stage retracts into the Echo liquid handler and the source plate gripper stage extends outside of the Echo liquid handler 7 Remove the source microplate and click the DONE button The source plate gripper stage retracts into the Echo liquid handler The liquid transfer protocol is finished Echo Liquid Handler Software 5 21 5 6 Labware window The LABWARE window is used to manage source and destination microplate definitions Figure 5 6 Labware window LA ET 555 Liquid Handler localhost oe x Protocols Labware Diagnostics Calibration Advanced LABCYTE gt Plate Name List 1536HB_DMSO 1536LD _Dest 1536LD _DMSO 1536LDY_DMSO_HT 384LD _AQ_B 384LD _AQ_P 384LDV_Dest 384PP_A4Q_BP 384PP_AQ_SP 384PP_Dest 384PP_DMSO 38LDY_DMSO Greiner_1536HiBasePS_782096 Greiner_384P5_761096 Add Edit Remove Plate Name List The PLATE NAME LIST contains all of the source and
31. coupling fluid pump is turned on the coupling fluid can flood the bottom of the system 6 8 Echo Liquid Handler User Manual 6 2 3 Clean and refill the coupling fluid bottle Clean and refill the fluid coupling bottle when one of the following occurs m Fluid appears cloudy m Two weeks have passed since the last coupling fluid refill m Fluid transfer accuracy or precision has been affected by coupling fluid contamination amp Note During periods of heavy duty cycle use refilling the coupling fluid bottle will be necessary more frequently than once every two weeks It is acceptable to skip the complete fluidic circuit purge as long as it is performed at least every second week The following procedures are needed to clean and refill the coupling fluid bottle 1 Empty the coupling fluid bottle 2 Purge the fluid circuit 3 Clean and refill the coupling fluid bottle 4 Cycle fluid through the Echo system This section describes each task in detail Tools and materials needed m 4 Liters of distilled water The Echo fluidic system holds about 1100 mL of fluid approximately 225 mL in the fluidic circuit and 875 mL in the coupling fluid bottle Emptying purging and filling the system will be performed several times A Caution Do not use de ionized water as it may corrode the fluidics system m An empty clean 1000 mL coupling fluid bottle square bottle catalog number ECHO AL15 S round bottle catalog numb
32. creates an analog RF radio frequency wave 2 The RF wave is amplified and transmitted to a focused acoustic transducer which transforms the RF wave into an acoustic pulse 3 This acoustic pulse is transmitted into the fluid in the source microplate well causing the fluid to form a droplet that travels upwards 4 The droplet is captured by the receiving surface which may be an fi inverted destination microplate well or a flat substrate Surface RF Pulse tension of the fluid keeps it on the receiving surface For more information about ADE Technology see Appendix A 1 1 1 How does the Echo system work The Echo liquid handler uses the following basic steps to transfer liquid 1 The operator loads the source microplate into the Echo liquid handler This step can also be performed by a robot D d The Echo liquid handler retracts the source microplate then surveys and measures the fluid height in each a well and the DMSO water Loading source microplate _ concentration for DMSO based solutions Echo 550 and Echo 555 only a DO Note The DMSO water concentration reported is not relevant for non DMSO solutions and should be ignored 2 The operator loads the microplate that will receive the liquid into the Echo liquid handler destination microplate Before retracting the destination microplate the Echo liquid handler inverts the microplate with the open microplate wells facing down
33. destination plates that have been defined for your system may vary from the list in the previous figure The source microplate types are specific to liquid types and use the naming convention platename_liquid type such as 384PP_DMSO Specialized features are appended to the plate name such as 1536_LDV_DMSO_HT This plate type is calibrated for high throughput HT dispensing which uses a faster transfer rate for higher volume or where precise droplet placement is not required See 1536 plate to 1536 plate liquid transfer on page 1 16 Since source microplates are carefully calibrated by Labcyte they cannot be added or removed by the user and only the barcode location can be edited The destination microplate types are specific to liquid types and use the naming convention platename_liquidtype_Dest such as 1536LDV_DMSO_Dest Destination plate types can be added edited or removed 5 28 Echo Liquid Handler User Manual O Note If you purchased an Omics package the Plate Name List will include the following entries m 384LDV_AQ B use for transfer of aqueous buffers such as PBS or TE with or without nucleic acids from 384 well LDV plates m 384LDV_AQ BP use for transfer of aqueous buffers containing protein including BSA from 384 well LDV plates m 384PP_AQ SP use for transfer of aqueous buffers containing surfactants such as Tween 20 or Triton X 100 from 384 well LDV plates m 384PP_AQ BP use
34. dialog box Change confirmation is not requested Apply The APPLY button displays a dialog box asking for confirmation of the AMPLIFIER FEEDBACK and or SYSTEM GAIN change s Answering YES applies the changes Answering NO closes the confirmation dialog box without applying the change s and returns to the CALIBRATE POWER dialog box Cancel The CANCEL button closes the CALIBRATE POWER dialog box without taking any further action 5 56 Echo Liquid Handler User Manual 5 8 3 Transducer calibration There are two acoustic transducer calibration tools m Focus calibration measures the focal Transducer length of the acoustic transducer and compares it to the previous Focal Sweep a measurement If no significant focal RES Les length change has occurred no recalibration is required and the focus calibration procedure is finished Focal Sweep is a diagnostic tool used by Labcyte personnel to measure the focal length of the acoustic transducer during a predefined vertical movement See Focal Sweep on page 5 59 Focus calibration The Echo acoustic transducer includes a lens that focuses the acoustic energy pulse at a specific point near the microplate well fluid surface Correct droplet ejection requires the ability to accurately focus the acoustic energy in the well fluid Changes to the transducer focal length over time may lead to a mispositioning of the Z axis Measurement of the DMSO water ratio during a source plate su
35. for transfer of aqueous buffers containing no surfactants from 384 well LDV plates Source plates require very exacting specifications and calibration to accurately transfer nanoliter volumes therefore they are defined specifically for the Echo instruments and tested at Labcyte Echo qualified For this reason source plates cannot be defined by the user For existing source plates only the barcode location can be edited see Figure 5 1 Contact Labcyte to add more source plates See Contact information on page 7 1 Figure 5 1 Plate Specification for Source Plate Plate Specification A1X Offset A 12 13 A1Y Offset 8 6 X Center Spacing C 45 mm Y Center Spacing D 4 5 mm Plate Height 144 mm Flange Height F 2 41 z mm Well width G 33 mm Well Capacity H 50 ul Source plate definitions cannot be removed from the Echo database or the LABWARE tab PLATE NAME LIST All source plate definitions can be used as destination plate definitions however no user defined destination plate definition can be used as a source plate definition Echo Liquid Handler Software 5 29 Add To create a new destination plate definition click the ADD button Edit To edit a plate definition either select the definition name and click the EDIT button or double click the plate definition name Source plate definition editing is limited to selecting a barcode location including none Destination plate d
36. from 1 src plate to 4 dest plates 2x2 or every other dest well 1536 well 1536 well standard 1 src plate to 1 dest plate Destacking transferring liquid from a single source src plate to several destination dest plates Stacking transferring liquid from several source plates to a single destination plate Examples of these protocols are described later in this section Additional and more complex fluid transfer protocols can be defined with the extensive Echo Activex control library as well as with the Echo applications software packages Echo Plate Reformat Echo Cherry Pick and Echo Dose Response These protocols can specify virtually any combination of transfers from source microplate wells to destination wells Refer to the Labcyte Echo Integration Guide for more information Liquid transfer can be tailored to the user s specific requirements Variables that are user selectable or definable include the following e Source microplate type from the list of Echo qualified plates e User definable destination microplate or surface e Transfer type for example interleaved or quadrant 384 well to 96 well transfers e Transfer volume e Number of copies how many times the transfer protocol is executed e Transfer map specifying exactly which wells or regions of wells will be transferred e Predefined DMSO water concentration maps These fluid transfer attributes and choices are defined in Chapter 5 i Note New fluid capabi
37. handler is both turned on and that its internal controller has completed its initialization and is ready 3 8 Echo Liquid Handler User Manual m Off When the Power Ready light is turned off either the Echo liquid handler has not been turned on or its internal controller has not completed initialization Warning light m Off Normally the Warning light is turned off indicating that the Echo liquid handler is functioning correctly Yellow The Warning light is turned on when any of the following conditions are true e The Echo liquid handler stages are not homed or are in the process of being homed See Home calibration on page 5 52 e The coupling fluid bottle level is low See Coupling fluid indicator on page 5 43 Note When the coupling fluid falls below its recommended minimum level air bubbles may be introduced into the coupling fluid which reduces the accuracy or precision of source plate surveys and fluid transfers Labcyte recommends that you refill the coupling fluid bottle as soon as possible after the Warning light has been turned on see Refill the coupling fluid bottle on page 6 7 e The coupling fluid temperature is outside of the normal range 22 1 C 0 9 C 21 2 C 23 0 C e The process door is neither fully open nor fully closed e The upper anti static bar is neither fully up nor fully down e The source plate dryer nozzle is neither fully up nor fully down Fault light m
38. it is reliable but also because very little maintenance is required m Routine operator maintenance takes only minutes each week typically keeping the coupling fluid bottle clean and filled m Field service maintenance is scheduled once every six months m The Echo touchless technology means that there are no tips no pins and no washing m The acoustic transducer itself has no moving parts to wear out It simply converts RF radio frequency energy to ultrasound energy The mechanisms that move it into position are designed for a long and serviceable life 1 2 6 Cost savings The Echo liquid handler reduces operating costs by m Enabling the routine use of higher density lower volume assays Moving to smaller assay volumes reduces the costs of compounds solvents and reagents m The use of higher density microplates High density microplates dramatically increase the number of assays that can be performed with a limited amount of test compound used m Minimizing the use of expendable supplies for example intermediate microplates tips pins and wash solutions m Eliminating washing stations and their complex procedures and fluids Introduction 1 9 1 2 7 Uncompromised accuracy precision and speed High throughput capabilities are maintained without degrading fluid transfer accuracy and precision Droplet to droplet volumetric precision exhibits a low CV coefficient of variation for all fluid types
39. method is to press CTRL A if at least one well is already selected Figure 5 2 Multiple regions defined in a transfer map Define Wells i Use your mouse to select the transfer wells To select more than one well use Control and Shift key combinations To select an entire row or column dick the appropriate header To select all wells click corner header or press ctrl A l SI ES Es amp am Click the OK button to save the transfer map definition with this protocol Echo Liquid Handler Software 5 23 5 5 2 Edit an existing protocol definition To edit an existing protocol definition click the EDIT button or double click the protocol name in the PROTOCOL List One of the following edit protocol dialog boxes will open Figure 5 3 Edit 384 384 protocol dialog box Edit Protocol Protocol Name 384 to 354 r Source Name 384PP_AQ_BP x Source 1 Wells AtiL6 View Define OK Figure 5 4 Edit 384 1536 protocol dialog box Destination Name Greiner_384PS_761096 ei Number of Copies FN Transfer Volume EN nt ps Edit Multi Plate Protocol Protocol Name 384 to 1536 r Source Name 284PP_AQ_5P ba Source 1 Wells ares View Define Source 2 Wells Aare View Define Source 3 Wells ECS View Define Source 4 Wells ECTS View Define Destination Name 1536HB_Dest Number of Copies 7 Transfer Volume 25 w E Transfer Map Type L Interleaved Quadrant Any
40. more than one well use Control and Shift key combinations To select an entire row or columr click the appropriate header To select all wells click corner header or press ctrl A B Det2 C D E C Det3 F G H C Dest 4 K ELLE C al L era lionel or AAE M ELEC km N a Er Gm SS lt BA Sa Sq aa a The Define Wells dialog box shows you the source wells white cells that can be transferred to the selected destination plate The gray cells represent the wells that are not available If you click the radio button for the different destination plates you will see that the transfer map changes for each destination plate Figure 5 5 Transfer maps for all four destination plates Use your mouse to select the transfer wells Use your mouse to select the transfer wells To select more than one well use Control and Shift key combinations To sel To select more than one well use Control and Shift key combinations To select an entire row or col click the appropriate header To select all wells click corner header or press click the appropriate header To select all wells click corner header or press ctrl 4 C Dest 1 E B Dest2 c D E C Det3 F G H C Dest4 DP PP N E Dest 4 1 K ff O Use your mouse to select the transfer wells To select more than one well use Control and Shift key combinations To se Use your mouse to select the transfer wells n To select more than one well use Control and Shift key c
41. precautions viii The Echo liquid handler has been designed for safe operation The safety warnings and precautions in this section and throughout the manual must be observed during installation relocation maintenance repair and normal operation of an Echo liquid handler Failure to comply with these warnings and precautions or with specific cautions and warnings found elsewhere in this manual violates the safety standards of design manufacture and intended use of an Echo liquid handler This can result in hazardous exposure to laser light high voltage or moving parts Exposure to these hazards can cause severe injury Safety notation marks This manual uses the following symbols in the left margin to draw your attention to the specified type of information Symbol Meaning A Warning Warnings alert all users to the following m Potentially hazardous conditions m Actions that may result in personal injury or death A Caution Cautions alert the user to actions that may result in the following m Damage to the equipment m Lost or corrupted data m Unrecoverable interruption of the operation being performed O Note Notes emphasize or expand upon the surrounding information Echo Liquid Handler User Manual Electrical safety warnings Always observe the following electrical safety warnings Warning Plug the Echo liquid handler into a grounded circuit Class 1 capable of delivering at least m 15A for
42. provides additional controls that may be useful to the experienced Echo user Figure 5 2 Advanced window Protocols Labware Diagnostics Calibration Advanced mTransfer Options Perform survey every D x plate s J Assume Homogeneous Plate DMSO F Power Monitoring IV Save Plate Survey Data to Disk M Save Transfer Data to Disk J Ignore Source Plate Sensor Check J Ignore Destination Plate Sensor Check r Solvent Concentration i Use predefined Define m Echo Liquid Handler Soft Reset Hard Reset reboot Shutdown 5 9 1 Advanced liquid transfer controls Each of these controls affects transferring fluids from a source plate to a destination plate They are global in scope and affect the execution of each liquid transfer protocol that is run Each setting has a default state enforced when the Echo software is launched or restarted User choices are not stored or remembered upon shutdown Survey interval Select the interval between source plate surveys by choosing a value from Perform survey every 1 plate s the PERFORM EVERY SURVEY drop down list The default setting of every 1 plate causes the source plate to be surveyed every time a new source or destination plate is loaded A survey can take anywhere from 60 seconds to 2 minutes A survey will always be performed when a new source plate is loaded Echo Liquid Handler Software 5 63 If the source plate fluid is stable
43. specified source microplate well into the corresponding destination microplate well A progress bar keeps you informed about the liquid transfer that is in progress If you need to abort the liquid transfer process click the CANCEL button 5 When the liquid transfer is complete click the NExT button Figure 4 13 Transfer in progress Run Protocol 384 10nL 5 copies 10 0nl 384 384 j Transferring plate 3 of 5 Source plate barcode Destination plate barcode COCCO COC E N Em lt Bach Hext gt The destination plate gripper stage extends outside of the Echo liquid handler and reverts the plate to its upright position 6 Remove the destination microplate and click the NEXT button Figure 4 14 Removing destination plate Run Protocol 384 10nL 5 copies 10 0nl 384 384 1 Please remove destination plate 5 of 5 from the gripper The destination plate gripper stage retracts into the Echo liquid handler and the source plate gripper stage extends outside of the Echo liquid handler Manual Operation 4 15 7 Remove the source microplate and click the DONE button Figure 4 15 Removing source plate Run Protocol 384 10nL 5 copies 10 0nl 384 384 The source plate gripper stage retracts into the Echo liquid handler The liquid transfer protocol is done 4 16 Echo Liquid Handler User Manual CHAPTER ECHO LIQUID HANDLER SOFTWARE The Labcyte Echo liquid handler comes with user softw
44. the Miscellaneous section there is a drop down list of miscelaneous diagnostic procedures and a LAUNCH button E Dry Plate Dry plate The DRY PLATE procedure vacuums the water film off the Maintenance Mode M Q bottom of a source plate If a source plate is not present the DRY PLATE procedure is aborted During the Dry PLATE procedure the acoustic i coupler nozzle is moved down and out of the way mt x and the coupling fluid pump is turned off J Dry Piate is completed successfully The plate dryer nozzle makes a pass to ensure that all of the moisture is removed then displays a message when it is done Move to well The Move To WELL procedure positions the acoustic transducer directly beneath a specified well 1 Select Move TO WELL and click the LAUNCH button The Move To WELL dialog box is displayed 2 Enter the row and column address of the well Example A1 Figure 5 1 Move to well function Move To Well Plate Type well ai Move To Cancel 3 Click the Move To button to move the acoustic transducer directly beneath the specified well If you want to abort this function click the CANCEL button 5 46 Echo Liquid Handler User Manual Diagnostic Survey The Survey procedure performs an acoustic survey of the fluid in the loaded source plate 1 Go to the DIAGNOSTICS window and use the Diagnostics controls to load a source plate into Survey gt the Echo liquid handler 2 Se
45. the width of the well at its bottom in millimeters This is not an SBS specified dimension It is used for source plates and is not necessary for destination plates Example For a 384PP_DMSO plate type G is 3 3 mm The valid range for G is 0 00 to 86 00 mm 5 34 Echo Liquid Handler User Manual H Well volume Dimension H specifies the volume of the well in microliters Example For a 384PP_DMSO 50 0 pL The valid range for H is greater than 0 0 pL L Note The Echo liquid handler does not currently use the destination well volume property and any well volume entry is ignored The OK button saves the new or edited definition The CANCEL button abandons the data that has been entered SBS standards The SBS Society for Biomolecular Sciences standards that may be useful include m ANSI SBS 1 2004 Footprint Dimensions m ANSI SBS 2 2004 Height Dimensions m ANSI SBS 3 2004 Bottom Outside Flange Dimensions m ANSI SBS 4 2004 Well Positions m SBS 5 Draft Side Wall Rigidity The SBS standards can be found at http www sbsonline org i4a pages index cfm pageid 3731 Echo Liquid Handler Software 5 35 5 7 Diagnostics window The DIAGNOSTICS window provides the following control and monitoring options m In the DEVICE CONTROL AND STATUS section the user can control many of the Echo internal devices and mechanisms The current position of those devices and mechanisms is reported by green lamp indicators Clicking the che
46. through the Echo system Its solid state design is compact environmentally friendly energy efficient and designed for long life and ease of use Your Labcyte service representative will set up the chiller with the Echo system Refer to this section for safety information equipment setup if the Echo system is relocated operation and troubleshooting Figure C 1 Labeyte chiller C 1 C 1 Shipping contents The Labcyte chiller is packaged with the following components m Labcyte recirculating liquid chiller m 100 watt 12 VDC table top power supply m AC line cord Chiller tubing is included in your Echo accessories C2 Safety Warnings and Precautions The Labcyte chiller has been designed for safe operation Observe all safety warnings and precautions in this section during installation relocation maintenance and normal operation of a chiller A Warnings Warning Do not use the chiller if the power cord is frayed This condition can cause a shock hazard Warning Do not disassemble the chiller m Access to internal components can cause a shock hazard m Improper servicing can damage the chiller unit and void the warranty A Cautions Caution Do not operate the chiller above 40 C Caution Do not store the chiller above 100 C Caution Do not obstruct the rear fan restriction of air flow will impair performance Maintain a minimum of 3 inch clearance around the rear fan C 2 Echo Liquid Handler User Manual C
47. to read the barcode on the destination plate after it is inverted Labcyte provides barcode scanners that support the following barcode types e Code 93 Code 39 Code 128 Codabar Pharmacode Interleaved 2 of 5 UPC EAN Contact Labcyte Service and Support to install or move the barcode scanners See Contact information on page 7 1 See Barcode Locations on page B 1 for limitations on where barcode labels can be placed on a microplate 3 12 Echo Liquid Handler User Manual 3 2 9 AC power and fuse compartment The AC power and fuse compartment as called the power input module is located on the rear panel to the left of the Emergency Off button It contains the main Echo liquid handler power on off switch AC power outlet and fuse compartment The Echo liquid handler instruments are built in two AC power configurations 230 VAC and 100 115 VAC The fuse compartment holds two fuses For information on replacing fuses see Replace the AC power fuse on page 6 17 Figure 3 1 AC power and fuse compartment Fuse Compartment Power On Off po 1 O Off On i oerni AC Power Outlet p System Description 3 13 Data connection The Echo liquid handler includes several communication connections to the right of the Emergency off button The RJ 45 Ethernet port is the only data connection required for normal Echo liquid handler operation The Ethernet cable connects the Echo liquid hand
48. volume to be transferred the composition of the fluid individual acoustic transducer performance and other factors The Echo liquid handler builds up larger volume transfers by ejecting multiple 2 5 nL droplets The multi droplet ejection from a single well will vary depending on the fluid composition and well fluid thickness height A 6 Echo Liquid Handler User Manual A 4 3 Transferring the droplet Figure A 4 illustrates the ADE components discussed above and shows a droplet being ejected from a 384 well to a 1536 well microplate The coupling fluid distilled water is pumped into the acoustic transducer assembly It exits through the coupler nozzle and flows down into the coupling fluid catch basin The coupling fluid is responsible for m Maintaining a stable acoustic transducer temperature Transducer performance is not consistent if its temperature changes during use m Efficiently coupling the acoustic energy between the transducer lens and the bottom of the source microplate If the acoustic energy were to travel through air it would be substantially attenuated and the Echo acoustic droplet ejection would not function correctly The coupling fluid is returned to the coupling fluid bottle via a suction created by the vacuum system Figure A 4 ADE diagram Transferred fluid Destination plate Ejected droplet gt Source plate Fluid being transferred Coupling fluid catch Gouplin
49. water has reached the empty waste bottle required level m The FAULT indicator on the front panel of the Echo liquid handler is red as a result of the full waste water bottle m Three months have passed since the bottle was emptied Use the following procedure to empty and clean the waste water bottle 1 Ensure that no fluid transfer or diagnostic procedures are under way 2 Disconnect the vacuum line from the fluidics panel to decrease vacuum pressure Figure 6 1 Diusconnecting vacuum 3 Unscrew the cap on the waste water bottle 500 mL bottle with red cap and discard the waste water 4 Clean the waste water bottle and tube a Add Alconox or hand soap and water to the bottle b Scrub the bottle and tube with a plastic bristle bottle brush c Thoroughly rinse the bottle and tube to ensure there is no soapy residue Use distilled water for the final rinse E Note If you see foam while refilling the bottle rinse the bottle and tube again with hot tap water 5 Inspect the bottle bottle cap and seal ring for cracks other damage or foreign material that would cause a vacuum leak Replace if necessary 6 Replace the cap to the waste water bottle Attach the cap securely to ensure a tight seal for vacuum hand tighten 7 Reconnect the vacuum line Maintenance and Service 6 13 6 2 5 Replace the water filter During plate survey and fluid transfer the coupler fluid is exposed to dust and particles from the ai
50. 072 7 717 544 European Patent EP 1337325 1324823 Additional patents pending in the United States and other countries END USER LICENSE AGREEMENTThe sale of the Products hereunder include a limited nontransferable nonexclusive license from Seller to Buyer under the above listed patents and any foreign counterparts thereto and other patents pending or owned by Seller solely to use the Products to practice liquid transfers and related processes described in such patents collectively Echo Technology is solely for the internal research and development activities of Buyer solely when the Echo Technology is used in conjunction with the Products Seller does not grant any right or license hereunder to and Buyer hereby agrees not to make any commercial use except as set forth above of the Echo Technology including without limitation use of the Echo Technology to i manufacture physical products for external sale ii provide diagnostic services or iii perform any of the foregoing on behalf of any third party Printed in the United States of America ii Echo Liquid Handler User Manual Contents Preface Intended audience vii Safety warnings and precautions 2 1 a viii Safety notation marks viii Electrical safety warnings ix Additional safety warnings X Safety cautions o oo ect mre aera Uae ct ans X Emergenc
51. 0O csv File Edit View Insert Format Tools Data Window Help Adobe PDF DSHS 46RY t BAST noaea 2 12 2004 Time 9 32 42 1 2 3 4 5 98 49 98 24 98 63 98 81 97 85 99 90 99 27 98 39 98 59 95 77 99 26 97 97 98 23 97 69 98 21 98 30 99 00 99 36 98 84 97 56 97 73 96 43 99 11 98 50 99 40 moor 1 10 Echo Liquid Handler User Manual 1 3 What types of liquid transfer are supported The Echo liquid handler supports a variety of simple to complex microplate liquid transfer protocols The bundled Echo software program allows a user to set up the following types of protocol transfers Table 1 1 Liquid transfer types Source Src Destination Transfer map type Microplate Dest Microplate full or partial 384 well 96 well destacking quadrant destacking interleaved 4 regions from 1 src plate to 4 dest plates 2x2 or every other dest well 384 well 384 well standard 1 src plate to 1 dest plate 384 well 1536 well stacking quadrant stacking interleaved 4 src plates to 4 regions on 1 dest plate 2x2 or every other dest well stacking quadrant stacking interleaved 384 well 3456 well 9 src plates to 9 regions on 1 dest plate _ 3x3 or every third dest well destacking quadrant destacking interleaved 1536 well 96 well 16 regions from 1 src plate to 16 dest plates 4x4 or every fourth dest well destacking quadrant destacking interleaved 1536 well 384 well 4 regions
52. 1 of 3 Protocol Name 1536 to 384 Source Plate Type 1536 0v_pmso Destination Plate Type SEEYN 3 Click the NEXT button The second ADD New PROTOCOL WIZARD dialog box opens Figure 5 2 Add new protocol wizard page 2 of 3 Transfer Map Type cy Interleaved Number of Copies fp E C Quadrant Transfer Volume 2 5 nL a Transfer Map Type Select an interleaved or a quadrant liquid transfer protocol type See What types of liquid transfer are supported on page 1 11 b Number of Copies Enter the number of times the protocol will be executed The valid range is 1 to 10 000 copies You are prompted to load and unload the source microplate and the specified number of destination microplates Transfer Volume Enter the transfer volume in 2 5 nL increments up to 2000 nL or up to 500 nL for aqueous transfers If you enter the transfer volume directly instead of using the increment decrement buttons remember that the value must be an even multiple of 2 5 nL 4 Click the NEXT button The third ADD NEw PROTOCOL Wizard dialog box opens Figure 5 3 Add new protocol wizard page 3 of 3 Source 1 Transfer Wells I View Define 5 20 Echo Liquid Handler User Manual 5 Click the View DEFINE button to create a transfer map for the 1536 well source plate The Define Wells dialog box opens Figure 5 4 Define wells template for interleaved plate map Define Wells Use your mouse to select the transfer wells To select
53. 4 8 buffer w protein 1 384LDV aqueous plate precision and accuracy are only guaranteed for a maximum depletion of 500 nL from a single well without resurveying The software will present a warning message when the depletion exceeds 500 nL but will allow you to continue Labcyte recommends that you test the performance of any single well depletion greater than 500 nL without a re survey Destination microplates All 96 384 and 1536 well plates that meet ANSI SBS 1 2004 standards for plate dimensions and are less than 16 mm in height Short and medium flange height microplates SBS 3 4 1 and 4 2 are supported Tall flange height microplates SBS 3 4 3 are not supported Supported microplate height SBS 2 4 2 1 1 is 8 0 16 mm 0 31 0 63 in Barcode types e Code 93 e Code 39 e Code 128 e Codabar e Pharmacode e Interleaved 2 of 5 e UPC EAN 3 3 9 Coupling fluid Fluid Distilled water algaecide Approved algaecide Labcyte catalog number ECHO AL04 4 mL bottle with dropper Contact Labcyte to reorder Water algaecide ratio 50 uL one drop algaecide per 1000 mL distilled water 2 ANSI American National Standards Institute SBS Society for Biomolecular Sciences For information on plate standards go to www sbsonline org msdc pdf ANSI SBS 1 2004 pdf 3 22 Echo Liquid Handler User Manual CHAPTER MANUAL OPERATION This chapter describes the manual operation of the Echo Liquid Handl
54. 4 well quadrant fluid transfer Wells from a single 1536 well plate can be transferred or destacked to four 384 well plates Individual wells or all wells from each quadrant can be transferred to the corresponding wells in the destination plate Figure 1 13 1536 to 384 quadrant full plate transfer 384 well 384 well Destination Microplate Destination Microplate 1 2 1536 Well Destination Microplate AF48 gt P24 384 well 384 well Destination Microplate Destination Microplate 3 4 Introduction 1 15 1 35 1536 plate to 1536 plate liquid transfer Fluid transfers between two 1536 well microplates are exactly the same as 384 plate to 384 plate transfers except for the well densities is Note Echo 555 only High throughput transfer of DMSO is available for the 1536 well LDV plate 1536LDV_DMSO_HT This special calibration type uses the same 1536 well LDV plate that is used for standard DMSO and aqueous transfer however the repetition rate for droplet transfer is much faster This calibration type is ideal for preparing multiple screening plates and prewetted plates CS Note For liquid transfer of very small volumes or transfer that requires precise q y q p droplet placement use the 1536LDV_DMSO calibration 1 16 Echo Liquid Handler User Manual 1 4 What system configurations are supported The Echo liquid handler can be used in a manual stand alone configuration or in several levels o
55. 6 Echo Liquid Handler User Manual 5 9 3 Advanced Echo control There are cases when the Echo controller must be shutdown or restarted The Echo controller is the hardware and software resident on the Echo instrument that controls all functions of the instrument Figure 5 2 Echo controller restart or shutdown Echo Liquid Handler Soft Reset Hard Reset reboot Shutdown A Caution If the Echo software is connected to a stub mode Echo liquid handler service running on the client PC clicking the HARD RESET button will reset the client PC and clicking the SHUTDOWN button will shutdown the client PC Soft Reset Clicking the Sort RESET button causes the Echo service to be restarted on the Echo controller Hard Reset Clicking the HARD RESET button causes the Echo controller to reboot After rebooting the Echo controller any attached client computer programs must be reconnected to the Echo controller Shutdown Clicking the SHUTDOWN button causes the Echo controller to perform its normal shutdown procedure If an Echo liquid handler needs to be turned off its controller should always be shutdown before the Echo AC power switch is turned off Echo Liquid Handler Software 5 67 5 68 Echo Liquid Handler User Manual CHAPTER MAINTENANCE AND SERVICE Daily maintenance consists of refilling the coupler fluid bottle checking for leaks in the instrument and positioning of the microplate insert Peri
56. 6 Small cell mode operation Ethernet Network O AAA aS Master Source Microplates cl U ey Destination Microplate Small Cell Reformatted Assay Microplates Controller Introduction 1 19 1 4 4 Fully automated operation A more fully automated operation expands on the small cell concept and may incorporate a larger range of processes and operations Figure 1 17 Full automated mode operation Larger fully automated drug discovery labs might include the following e A larger combination of the processes and Stations listed in the discussion of a small cell mode operation See Small cell operation on page 1 19 e One or more supervisory workstations e A conveyor system to move microplates and other material not only between stations but into and out of the lab e Multiple robots individually chosen and programmed to support a specific function or machine e Multiple Echo liquid handlers Echo Liquid Handler iii Microplates D Note Contact your Labcyte field engineer to assign different IP addresses to your Echo devices Echo Liquid Handler SIT wa om a o a a a aa oa oa oa oa a aa piolniot LITITIIIIIIIIT1 S _ 1 20 Echo Liquid Handler User Manual 1 45 Multi client considerations Larger research labs may have many computers instruments controllers and other dev
57. 61 2 Click NEXT to display the BARCODE SCANNER CALIBRATION WIZARD dialog box Figure 5 1 Barcode scanner calibration wizard dialog box Barcode Scanner Calibration Wizard Page 2 of 6 Scanner Configuration Please choose the scanner configuration for your system L A The barcode used to calibrate the scanner is at Source Plate Left Side Destination Plate Clicking Next will open the door and present the gripper us LES re The selections available not grayed out depend on the barcode scanners installed in an Echo liquid handler 3 Select the barcode scanner configuration you want to calibrate plate type and barcode and click NEXT The selected source or destination plate gripper stage will be extended outside of the Echo liquid handler 4 When prompted load a microplate that has an appropriate barcode label 5 Click NEXT to begin the calibration The microplate is retracted into the Echo liquid handler where the barcode scanner calibration is performed 6 When the calibration is complete click NEXT to extend the source or destination plate gripper stage outside of the Echo liquid handler 7 Remove the plate and do one of the following m Click the NExT button to perform another barcode scanner calibration or m Click the FINISH button to exit the barcode scanner calibration wizard 5 62 Echo Liquid Handler User Manual 5 9 Advanced window The ADVANCED window
58. 7 1 2 Finish any operation in progress 3 Remove any microplate from the Echo liquid handler 4 Turn off the Echo liquid handler and the fluid chiller See Turn off Echo power on page 4 4 and Chiller Setup and Operation on page C 3 5 After the Echo liquid handler has been powered down disconnect the following lines from the back panel Chiller input and output tubing Compressed air supply line Vacuum supply line Ethernet cable AC power cable Coupling fluid bottle caddy and cable 6 Use proper safety procedures to lift the Echo liquid handler gently onto a moving dolly or cart of sufficient size and strength to support the unit A Warning The Echo liquid handler weighs approximately 128 kg 283 Ib Moving or lifting the Echo liquid handler incorrectly can cause severe injury Never attempt to move or lift the Echo liquid handler without using proper equipment and trained personnel 7 Move the Echo liquid handler and ancillary equipment and material bottles tubing microplates source plate inserts etc to the new location which should already be prepared and comply with all pre installation requirements See Pre installation requirements on page 2 2 2 12 Echo Liquid Handler User Manual 2 4 2 Reinstall the Echo liquid handler after a move After the move use the following procedure to restore an Echo liquid handler to normal operation 1 Ensure that the new mounting surface is le
59. 92 168 1 1 and a subnet mask of 255 255 0 0 Check with your System or Network Administrator to make sure this address is available Figure 2 1 Entering host name i Echo 5XX Labcyte Inc Xl Welcome to the InstallShield Wizard for Echo 5XX The InstallShield R Wizard will install Echo 5XX on your computer To continue click Next Install Client Only f Stub Mode LABCYTE smstatserver only E NormallMode gt C Install Client Server Please enter remote host name or IP address 555 0758 WARNING This program is protected by copyright law and international treaties After the installation type has been selected click NEXT 3 Read the release notes They contain information about the version of software being installed The release notes will also be stored in the Labcyte Echo liquid handler folder on your Windows Desktop for later viewing Click NEXT Installation and Relocation 2 1 Figure 2 2 Echo software release notes fig Echo 5XX InstallShield Wizard Eg Read Me Please read the Following instructions before installing the software Release Notes for the Echo Liquid Handler Software v2 3 0 z Copyright c 2003 2010 Labcyte Inc All Rights Reserved Version Release Date September 6 2010 Contents FOR MORE INFORMATION INSTALLATION Office 2003 COMPATIBILITY WARNING WHAT S NEW Echo Liquid Handler RELEASE NOTES AND KNOWN ISSUES Installshiel
60. Aqueous buff protein to 1536LDY 5 copies 2 5nl 384 1536 I 384PP DMSO to 384P5 10 copies 5 0nl 384 384 Add Edit Remove The ABouT window displays the current software version and build numbers and displays the copyright information Figure 5 3 About window About Echo Liquid Handler LABCYTE gt Version 2 3 0 Build 20100913 103 Copyright Labcyte Inc 2003 2010 All Rights Reserved 5 6 Echo Liquid Handler User Manual 5 3 2 Help window You can display the HELP window by right clicking on any tabbed window Protocols Labware Diagnostics Calibration or Advanced and selecting Help in the menu You can also open the HELP window by pressing the F1 key The HELP window displays the page that corresponds to the function window you have currently open The following figure shows the help page for Protocols Figure 5 4 Help window E Echo Liquid Handler Client Help A Home a Print Options Navigation St 4 Back options Contents index Search Z Introduction B Multi client considerations Protocols gt Labware gt Diagnostics gt Calibration Content Index and Search tabs Help topic Help page Links to related subtopics To use the HELP window O x Echo Liquid Handler Help Protocols The Protocols tab is the first window that opens when you start the Echo liquid handler software The Prot
61. CS Note Only Echo qualified source microplates are listed in the Source Plate Type drop down list Contact Labcyte to add more source plates See Contact information on page 7 1 Destination Plate Type Select a destination plate type from the second drop down list The destination plate definition must already exist in the Echo database Figure 5 1 Add new protocol wizard page 1 of 2 Add New Protocol Wizard Protocol Name 384 10nl Source Plate Type 384PP_AQ_BP v Destination Plate Type On Echo Liquid Handler Software 5 13 2 Click the NEXT button The second ADD New PROTOCOL WIZARD dialog box opens Figure 5 2 Add new protocol wizard page 2 of 2 Add New Protocol Wizard Page 2 of 2 Transfer Wells View Define Number of Copies 1 m Transfer Volume 2 5 nL 3 Transfer Wells Click the ViEW DEFINE button to select the wells to be transferred from the source plate a Select the source plate wells that contain the compound to be transferred See Figure 5 3 For information on how to select transfer wells see Create a transfer map on page 5 22 b Click the OK button to return to the previous screen Figure 5 3 The Define Wells dialog box Define Wells e Use your mouse to select the transfer wells To select more than one well use Control and Shift key combinations To select an entire row or column dick the appropriate header To select all wells click corner header or press ctr
62. Contact information on page 7 1 2 4 Echo Liquid Handler User Manual 2 2 Installation overview The Labcyte Echo liquid handler is installed by Labcyte field engineers The following is a brief overview of the hardware and software installation steps they perform 1 7 8 Unpack the Echo liquid handler and place it in the area that meets the minimum operating space requirement Connect the Echo liquid handler to AC power air pressure vacuum pressure surge tank optional coupling fluid bottle and the coupling fluid chiller Fill the coupling fluid bottle and prime the coupling fluid circuit Perform a basic functional check of the Echo liquid handler Perform the Site Acceptance Test SAT process optional Change the Echo IP address if a different IP address is required by your system administrator Install the Echo software onto the customer supplied PC client workstation Connect the client PC to the Echo liquid handler and verify communications Figure 2 1 Installed system Ethernet cable Le cross over e Compressed air line Vacuum line DU teen 533 led CCC Echo liquid handler Chiller fluid lines Client PC Bench Floor ra s me Surge tank optional Chiller Installation and Relocation 2 5 2 3 Software installation Labcyte installs the Echo software onto the client PC that is connected to the Echo liquid handler If you need to install the softwa
63. Echo software window is a pair of message log windows server and client and a set of checkboxes that select which messages are logged Figure 5 5 Log windows and controls 09 24 03 07 27 13 841 FluidTransferServer cpp 2953 DEBUG Get4llsrcPlateNames call 09 24 03 07 27 14 841 FluidTransferServer cpp 2996 DEBUG MoveZ ell called Server IV all N Debug M Warning M Info IV Error Client log messages Messages in the client log window originate in the client PC In addition to being displayed the client messages are stored on the client PC The exact location depends on the Echo software installation The default location for the client log files is C Program Files Labcyte Medman Data The most recent client log file is named client_localhost_MedmanGUI txt and older client log files are named client_localhost_MedmanGUI old 0 txt client 1 txt client 2 txt and so on The most recent ten historical client log files are stored in addition to the log file for the current session Server log messages Messages in the server log window originate in the Echo liquid handler Server log messages are stored in log files in the Echo controller CS Note During normal operation you do not need to know this information However if you are troubleshooting a problem your Labcyte field engineer may ask for this information 5 8 Echo Liquid Handler User Manual Select log message types Which types of data will appe
64. F energy pulses Power calibration is performed automatically by the Echo liquid handler as part of each liquid transfer protocol CS Note Normally a manual power calibration will not need to be performed by a user However it can be useful during certain repair diagnostic or recovery procedures Perform power calibration only as directed by Labcyte personnel If manual power calibration is required use the following procedure 1 On the CALIBRATION tab click the CALIBRATE button in the POWER section The CALIBRATE POWER dialog box will open Figure 5 1 Calibrate Power dialog box reporting current values Calibrate Power Reference Amp 1 6 volts Pulse Energy 3 mJ Power Amplifier Feedback 1 04069 Pulse Energy mJ Vpp volts Calibrate r System Gain 1 l Override Cancel Apply Reference m Amp The Amp field displays the reference tone burst amplitude m Pulse Energy The PULSE ENERGY field displays the reference pulse energy value 5 54 Echo Liquid Handler User Manual Power Amplifier Feedback The AMPLIFIER FEEDBACK field displays the current power correction factor and normally should be very close to 1 0 Pulse Energy The PULSE ENERGY field will be empty until power calibration is performed Vpp The voltage peak to peak VPP field will be empty until power calibration is performed 2 Click the CALIBRATE button in the POWER section of the CALIBRATE
65. Plate V In M out Q Present Destination Plate V In M out O Present Flip Destination Plate T up O Down Plate Dryer Up O Down Plate Dryer I On O Vor Anti staticBars On O Vor Upper Anti static Bar V up M Down O Coupler Nozze up Q V Down Coupler Pump On Q Vor Coupler Pump Direction V Normal Air Pressure Warning VacuumPressure coupiingFluid EMO 2 Waste Bottle Fluid Leak Focus Calibration Fluid Temp 21 97 C 43 45 C Dry Plate M Launch Maintenance Mode I Q 08 24 06 15 56 56 677 Client 127 0 0 1 localhost has connected to the Echo server 08 24 06 15 58 31 373 Null axis set Z position to nominal 16000 PlateSurveyB 08 24 06 15 58 31 451 Starting Plate Survey well A1 rows 16 cols 24 Plat 08 24 06 15 58 31 498 Analyzing A1A2A3A4ASA6ATABA9A10A11A12413A14A154 08 24 06 15 58 34 498 Analyzing B1B2B3B4B556B7B8B9510811B12513514B15B16I 08 24 06 15 58 37 499 Analyzing C1C2C3C4CSC6C7C8C9C10C11012013C14C15C16 08 24 06 15 58 40 515 Analyzing D1D2D3D4D5D6D7D8D9D10D11D12D13D14D15D16I 08 24 06 15 58 43 499 Analyzing E1E2E3E4ESE6E7E8E9E10E11E12E13E14E15E16E17 08 24 06 15 58 46 500 Analyzing F1F2F3F4FSF6F7F8F9F10F11F12F13F14F15F16F17 7 b Client FAIT Debug V Warning IV Info IV Error Version 2 1_Beta Build 00 060823 00 Copyright Labcyte Inc 2003 2006 All Rights Reserved Maintenance and Service
66. Processed homogeneous Processed inhomogeneous Ignore Source Plate Sensor GO CS Note Selecting processed or raw fluid percentages affects the following data m The data displayed in the plate map m The data displayed in the Statistics section m The data saved to a CSV file on the client PC The source microplate survey data is displayed in the main data grid of the SURVEY dialog box Either fluid composition or well fluid thickness height will be displayed depending on which RESULT radio button is selected The data will either be raw processed homogeneous or processed inhomogeneous depending on which Post PROCESSING radio button is selected Echo Liquid Handler Software 5 49 In the STATISTICS section the minimum maximum Statistics average and CV data from the survey is reported Mn 2012 These statistics are calculated from raw processed Max 2 149 homogeneous or processed inhomogeneous Avg 2 06221 survey data depending on which POST PROCESSING CV _29007 radio button is selected In the RESULTS section raw data collected during the survey is displayed This information is used only by Labcyte personnel during troubleshooting or system analysis Figure 5 1 Diagnostic survey results Results FW FW Well BB BB TB1 TB1 TB2 TB2 pe Pi D Lire ToF Vpp ToF Vpp ToF Vpp B2 1 826 3 2 15 0 984 2 5 132 2 053 77 7 B3 1 826 3
67. Proxy cpp 623 GetAllProtocolHame SOAPFluidTransferProxy cpp 131 Getinstrumentinfo Displ ay option S for 07 2240 14 23 57 870 SOAPFluidTransferProxy cpp 623 Getinstrumentinfo P PRE server and client status window gt Server ii Debug V warning IV Info M Error Version 2 3 0 Build 20100628 86 Copyright Labcyte Inc 2003 2009 All Rights Reserved 5 4 Echo Liquid Handler User Manual Tabs Each tab opens a window for a function or related group of functions e Protocols In the Echo software a protocol is a set of user selected parameters plate fluid type plate mapping transfer volume and repetition that define a single liquid transfer procedure The PROTOCOLS window lists the protocols available to run From this window you can create modify run or delete a protocol e Labware The LABWARE window shows a list of pre defined types of microplates You can select these microplates during the creation of a protocol From this window you can also create additional destination microplates e Diagnostics The DIAGNOSTICS window displays the status of the Echo liquid handler From this window you can also manipulate some of the individual motor controls and perform some maintenance tasks e Calibration The CALIBRATION window provides calibration procedures for microplate registration power modulation transducer focus and barcode alignment e Advanced The ADVANCED window provides additional liquid transfer op
68. RAMS 4 Select Echo liquid handler and click REMOVE 2 3 3 Upgrade the Echo software Upgrading the software requires a new Echo software CD Contact Labcyte Service and Support for the most current version See Contact information on page 7 1 Depending on your current software version you may need to run a clean installation Check with Labcyte Service and Support to determine if your software can be upgraded If not you will need to perform a new or clean installation Back up your existing liquid transfer and plate definition data 1 Insert the new Echo liquid handler software CD in the client PC 2 Follow the installation instructions See Install the software on page 2 7 Installation and Relocation 2 11 2 4 Instrument Relocation DO Note If you do not plan to set up the Echo liquid handler within two weeks refer to Extended Non Use and Storage on page 6 18 2 4 1 Prepare the Echo liquid handler for a move Use the following procedure if the Echo liquid handler must be moved 1 Determine if the move must involve Labcyte m f the move requires transportation truck air train call Labcyte Service and Support Do not attempt to move the Echo liquid handler yourself m Ifthe move can be easily accomplished using a moving dolly or cart the move can be performed without Labcyte involvement If you have any questions contact Labcyte Service and Support See Contact information on page
69. T P N 001 3966 230 VAC 50 Hz 5A 250 VAC 5 A FAST P N 001 3967 1 Turn off the AC power switch if it is not already off 2 Unplug the AC cord from the Echo power entry module 3 Pry open the power entry module fuse cover with a small screwdriver 4 Extract the fuse holder with a small screwdriver 5 Replace the two AC power fuses See Table 6 3 Figure 6 2 Replacing the AC power fuses Pry here to open cover Fuse Holder AC Power b Fuse Cover Fuse one of Entry Module two shown 6 Return the fuse holder to the power entry module 7 Close the fuse cover 8 Plug in the power cable 9 Turn on the Echo liquid handler Maintenance and Service 6 17 6 3 Extended Non Use and Storage The Echo liquid handler is designed to be used on a regular basis daily or within a week Regular use keeps the acoustic transducer wet and provides consistent liquid transfer results The acoustic transducer must remain wet to retain the high degree of precision and accuracy that is required for nanoliter volume transfer If the transducer is allowed to dry out the focal length can change which can affect DMSO measure and transfer accuracy If you do not plan to use the Echo liquid handler for longer than two weeks perform one of the following options m If the Echo liquid handler remains connected to the power source chiller air and vacuum lines then perform the maintenance procedure described in section 6 3 1
70. The acoustic echo ToF returned from the interface of the well fluid meniscus and the air above it m EW BB Empty Well Bottom of the Bottom The acoustic echo ToF returned from the interface of the bottom surface of the source plate bottom and the coupling fluid when the well is empty 5 60 Echo Liquid Handler User Manual m EW TB Empty Well Top of the Bottom The acoustic echo ToF returned from the interface of the top surface of the source plate bottom and the air in an empty well 6 If the data measured by the focal sweep procedure needs to be saved for analysis select FILE SAVE in the upper left corner of the focal sweep window 5 8 4 Barcode scanner calibration An Echo liquid handler may have optional microplate barcode scanners installed Several barcode scanner functions can be recalibrated If barcode scanning errors are experienced perform the following procedure 1 Click the CALIBRATE button in the BARCODE SCANNER section of the Calibration window The first screen in the Barcode Scanner Calibration Wizard opens Figure 5 2 Barcode scanner calibration wizard Barcode Scanner Calibration Wizard Page 1 of 6 MY UAU WA Welcome to the Barcode Scanner Calibration Wizard Cancel This procedure can be aborted by clicking the CANCEL button CS Note If an error occurs during this procedure an error message will appear and contain directions on how to proceed Echo Liquid Handler Software 5
71. Type bsap best M Dest 4 10 Echo Liquid Handler User Manual 3 Click the Next button The second window will open Figure 4 7 Add new protocol wizard page 2 6 Add New Protocol Wizard Page 2 of 2 Transfer Wells View Define Number of Copies 5 Transfer Volume 10 nt m lt Back Finish cane Enter the number of copies for example 5 Each copy represents a single plate transfer Enter the liquid transfer volume for example 10 nL If you enter the transfer volume directly instead of using the increment decrement buttons remember that the value must be an even multiple of the minimum transfer volume 2 5 nL If you enter any other value you will see an error message Click the ViEW DEFINE button to display the DEFINE WELLS window Figure 4 8 Defining transfer wells LI x Use your mouse to select the transfer wells To select more than one well use Control and Shift key combinations To select an entire row or column dick the appropriate header To select all wells dick corner header or press ctrl A 7 Specify the source wells from which to transfer fluid To define the region A1 through E4 click cell A1 and drag the mouse pointer to cell E4 8 Click the OK button to return to the previous window 9 Click the FINISH button to store your new protocol definition For more information about creating a 384 well protocol see Create a new 384 384 protocol definition on pa
72. ab is yellow indicating that the coupling fluid has dropped to the refill required level See Coupling fluid indicator on page 5 43 m LCD panel WARNING indicator has turned yellow as a result of the low coupling fluid level See Fault light on page 3 9 m Echo ActiveX events report that the coupling fluid has dropped to the refill required level Refer to the Labcyte Echo Integration Manual Materials required m Distilled water sufficient to fill the 1L bottle to 900 mL A Caution Do not use de ionized water as it may corrode the fluidics system m 50 uL one drop algaecide Labcyte catalog number ECHO AL04 m Gloves for protection when handling the algaecide m Paper towels or shop cloths to deal with any unexpected fluid leaks 1 Ensure that no fluid transfer or diagnostic procedures are under way 2 Turn off the vacuum and coupling fluid pump a Turn off the coupling fluid pump through the DIAGNosTIcs window of the Echo software b Unplug the vacuum line to decrease vacuum pressure CS Note The surge tank will add a few seconds to stopping the vacuum 3 Unscrew the cap on the coupling fluid bottle and remove the bottle from its caddy Be careful not to lift the coupler fluid tube straight tube above the water level or you will introduce air into the line If you have air in the line you will need to purge then recycle the fluid through the system for at least 15 minutes See Purge the fluid circuit
73. ar in the log windows can be specified by selecting one or more of the checkboxes at the bottom of the Echo software window All All message types debug warning info and error are displayed in the client and server log windows Debug Debug messages are generally not of use to Echo users They provide detailed information to Labcyte representatives Warning Warning messages report unexpected events These events may not cause a functional or operational problem since the Echo liquid handler can recover from most situations that create a warning Info Info messages report the normal activity of the Echo liquid handler Error Error messages are typically serious They may be triggered by events that cause an operation the Echo liquid handler or the Echo software to fail Echo Liquid Handler Software 5 9 5 5 Protocols window The PROTOCOLS tab is the first window that opens when you start the Echo liquid handler software The Protocol List displays the liquid transfer protocols that are stored in the Echo database A tool tip displays many of the protocol parameters From the PROTOCOLS window you can perform the following tasks m ADD create a new liquid transfer protocol definition EDIT change an existing protocol definition REMOVE delete an existing protocol definition RUN execute a liquid transfer protocol Figure 5 1 The Protocols window LA Echo 555 Liquid Handler localhost Protocols Labware Diagnosti
74. are The Echo liquid handler software referred to as the Echo software from this point on provides a Windows based graphical user interface GUI The user can control the Echo liquid handler and execute most of its functions which include the following Launch the Echo software start the Echo software Window description learn the Echo software windows and functions About and Help windows view the software version and online help Status window monitor Echo liquid handler functions Protocols window define new liquid transfer protocols and edit or delete existing protocols run the protocols Labware window define new destination plates and edit or delete existing destination plate definitions Diagnostics window monitor Echo liquid handler status directly control most of the Echo liquid handler motor function and perform diagnostic procedures Calibration window perform calibration procedures Advanced window Change transfer options reset or shut down the Echo controller Echo Liquid Handler Software 5 1 5 1 Launch the Echo software The Echo software can be launched by either of the following methods m Open the Labcyte Echo folder on the desktop and double click the icon for your Echo liquid handler Figure 5 1 Starting the Echo liquid handler from the Windows Desktop Labcyte Echo 5XX m Go to START gt PROGRAMS gt LABCYTE gt Echo 55X and select Echo Liquid Handler Software
75. as die ta das ae a AS ero 2 A 3 19 3 39 hent PE Anh Gna ye hon ag gee lh we ee Gh are Go aed 3 19 3 3 6 Communications tas a os vis See ob te eS 3 20 iv Echo Liquid Handler User Manual 4 1 4 2 4 3 4 4 5 1 5 2 5 3 5 4 5 5 5 6 5 7 5 8 5 9 3 3 7 Fluid transfer LT aa a a 3 20 3 3 8 Supported labware 3 21 3 3 9 Co pling ide ire amis Oe wee wee haut 3 22 Chapter 4 Manual Operation Startup Shutdown Le ee a 4 2 4 1 1 Turn on Echo POWs sa ey Jee NS RUES ESS 4 2 4 1 2 Turn off Echo power 4 4 Deline Labware 45 5 5 Se Rat oies WH Sb hos 4 7 4 2 1 Edit the source microplate 4 8 4 2 2 Define a new destination microplate 4 9 Create liquid transfer protocol 4 10 4 3 1 Add new POIDS Lena Led A una ue Ae at 4 10 Run the liquid transfer protocol 4 12 4 4 1 Prepare the microplates 4 12 4 4 2 Start liquid transfer run aaa a 4 13 Chapter 5 Echo Liquid Handler Software Launch the Echo software 5 2 Window description rar ar ua de te SCA NRC 5 4 About and Help windows 5 6 5 3 1 About window cys esc tact envi EN TENTE SN amp 5 6 5 3 2 Help window ieee seed ee oe ee Ener RA aR RAO Re 5 7 Status LL CN o oo aye e E a EE DIE ee 5 8 Protocols window se atk hei ad eh eh te re Naa a he al ee nr 5 10 5 5 1 Create
76. been activated See EMO indicator on page 5 42 m A coupling fluid leak caused by accumulation in the bottom pan has been detected See Fluid leak indicator on page 5 42 EMO indicator There are two EMO Emergency Motion Off switches one on the front and one on the rear of the Echo liquid handler Gray OK The EMO switches are in their normal not activated position Red An EMO switch has been activated Power has been turned off to many internal Echo electrical devices For a full discussion of the EMO switch when it should be used and how to power on an Echo liquid handler after the EMO switch has been used see EMO switch on page 3 10 Fluid leak indicator Sensors inside the Echo liquid handler monitor the coupling fluid system for leaks or unsafe fluid levels Green No coupling fluid leak or pending catch basin overflow has been detected Red Either a coupling fluid leak has been detected or the coupling fluid level in the acoustic transducer catch basin has risen to an unsafe level The coupling fluid pump has been turned off The user should call Labcyte Technical Support 5 42 Echo Liquid Handler User Manual Compressed air pressure indicator A pressure sensor inside the Echo liquid handler monitors the user supplied compressed air supply Green OK Air pressure is within specifications Red The user supplied compressed air is low lt 552 kPa lt 80 psi or nonexistent Check for leaks dis
77. bottle bottle cap and seal ring for cracks other damage or foreign material that would cause a vacuum leak Replace if necessary 3 Fill the coupling fluid bottle with 900 mL of distilled water and add 1 drop 50 uL of algaecide L Note The ratio of algaecide to water is optimized for deterring the growth of algae while minimizing residual moisture on the bottom of the plate 4 Return the cap and its attached tubing to the coupling fluid bottle 5 Attach the cap securely to ensure a tight seal for vacuum hand tighten and return the refilled coupling fluid bottle to the bottle caddy 6 Reconnect the vacuum line to the Fluidics Interface Panel A Caution Make sure that the vacuum line is connected and working correctly before turning the coupling fluid pump back on If the vacuum line is disconnected or the vacuum system is not operating correctly when the coupling fluid pump is turned on the coupling fluid can flood the bottom of the system Maintenance and Service 6 11 Cycle fluid through the Echo system A Caution The chiller does not need to be running for this procedure however do not cycle coupling fluid through the Echo liquid handler that is below 22 C or above ambient room temperature Extreme temperatures can damage the acoustic transducer 1 Turn on the coupling fluid pump The coupling fluid will be pumped into the Echo liquid handler After three to five minutes verify that the coupling fluid is r
78. ceive the fluid droplets ejected upwards from the source plate wells Figure 3 4 Inverting the destination plate Destination microplate up Plate being inverted Destination microplate down As the destination plate gripper stage retracts into the Echo liquid handler a sensor detects whether a plate is or is not present Normally a transfer will not be executed if a destination plate is not detected The human operator or a controlling program can override the plate presence check if necessary When the destination plate gripper stage extends out of the Echo liquid handler the plate is rotated back to its upright position 3 2 5 Status indicator lights There are three status indicator lights on the front of the Echo liquid handler and a duplicate set on the rear panel A quick glance can confirm whether the Echo liquid handler is operating normally or that it needs operator attention The Power light green is on when the instrument on and initialized the Warning light yellow turns on when a condition is outside of normal limits but the instrument can continue to operate the Fault light red turns on when a condition exceeds warning conditions and the instrument can no longer operate Figure 3 5 Echo liquid handler front panel Power Green Ready Warning Clear OK Yellow Warning Fault Clear OK Red Fault Power light Green Normally the Power Ready light is turned on indicating that the Echo liquid
79. cessive moisture gt 80 relative humidity Moisture or condensation can damage electrical components Caution Do not expose the Echo liquid handler to temperatures outside of the storage range 5 C 45 C 41 F 113 F Temperatures outside of this range can damage the instrument Caution Do not run the Echo liquid handler without the chiller Ensure that the chiller is correctly connected to the Echo instrument and is running at the recommended temperature Incorrect coupling fluid temperature can reduce the precision and accuracy of the fluid that is transferred Extreme fluid temperatures can potentially damage the acoustic transducer Caution Do not allow the acoustic transducer to dry out If the Echo liquid handler is going to be shut down for longer than a week start up and run the pump once a week to keep the acoustic transducer wet See Extended Non Use and Storage on page 6 18 Caution Do not run the pump if the chiller tubing is not connected Running the pump without the chiller tubing may damage other system components Caution Do not run the pump without the vacuum source connected and active Running the pump without vacuum can lead to flooding the system or a system error Preface xi Emergency motion off switch EMO The Echo liquid handler includes two EMO Emergency Motion Off switches They are located on the front and rear of the unit and are prominently labeled EMERGENCY OFF See P
80. ching one of the two following power configurations e 115 VAC 50 60 Hz 10A e 230 VAC 50 60 Hz 5 A Note The power configuration is factory set and cannot be changed onsite Compressed air source Clean dry air at 552 kPa 80 PSI minimum 1034 kPa 150 PSI maximum Consumption not to exceed 1cf m 150 PSI Labcyte provides the compressed air line and fitting with the Echo liquid handler but the customer must provide an LC series coupling body in line ferruleless with shutoff valve for 170 in ID 250 in OD tubing Vacuum source Vacuum returns the coupling fluid to the supply bottle and dries source plates before they are unloaded Customer must provide either house vacuum or vacuum pump with the following specifications e House vacuum Minimum vacuum at Echo connection 200 Torr 266 mbar 22 in Hg Vac Vacuum system capacity steady state 15 L min 0 9 m h 0 53 cf m Drying surge 80 L min for 10 sec 4 8 m h 2 8 cf m 33 max duty cycle e Vacuum pump Minimum vacuum at Echo connection 100 Torr 133 mbar 26 in Hg Vac Pump speed with 10 L surge tank 40 L min 2 4 m h 1 4 cf m without surge tank 80 L min 4 8 m h 2 8 cf m Note If the vacuum measured at the Echo connection dips below 1 2 an atmosphere 380 Torr 15 in Hg Vac then the Echo liquid handler could go into an error state To ensure consistent vacuum Labcyte recommends the use of a 10 L surge tank contact your Labcyte representative Ins
81. ck box next to the device initiates the action for example clicking Source Plate In directs the Echo liquid handler to retract the source plate gripper stage m In the SYSTEM STATUS section the user can monitor the current overall status of the Echo liquid handler through ten additional status indicators and two internal Echo temperature readouts m in the MISCELLANEOUS section the user can either activate the Echo maintenance mode or execute diagnostic procedures Figure 5 1 Diagnostics tab Lamp indicators LA ET 5XX Liquid Handler localhost Protocols Labware Diagnostics Calibration Advanced Device control and status Door V Open M Close Q Source Plate V in M out QO Present Destination Plate JV In F Out Present Flip Destination Plate Up O V Down Plate Dryer Up O V Down Plate Dryer FT On QO vor anti static Bars On O Vv of Upper Anti staticBar V Up M Down Q Coupler Nozzle up Q V Down Coupler Pump V On M off Oo Coupler Pump Direction JY Normal J Reverse Q System Status m Miscellaneous Power Air Pressure 9 Warning VacuumPressure ory Plate Zl Fault Coupling Fluid EMO E ee ates Launch Fluid Leak Focus Calibration AU AL O Coupling Fluid Temp 21 97 C RF Temp 1 43 45 C 5 36 Echo Liquid Handler User Manual 5 7 1 Device control You have access to precise Echo device control The ability to command internal mechan
82. connections or for the air pressure turned off at the source Vacuum supply pressure indicator A pressure sensor monitors the user supplied vacuum supply line pressure inside the Echo liquid handler Green OK Vacuum pressure is within specifications Red The user supplied vacuum pressure is low lt 78 kPa lt 15 in Hg Vac or nonexistent Check for leaks disconnections or for the vacuum turned off at the source Coupling fluid indicator A sensor monitors the level of coupling fluid in its bottle Green OK Coupling fluid is above the minimum required level Yellow The coupling fluid bottle level is low lt 250 mL LS Note When the coupling fluid falls below its recommended minimum level bubbles may be introduced into the coupling fluid reducing the accuracy or precision of source plate surveys and liquid transfers Labcyte recommends refilling the coupling fluid bottle as soon as possible see Refill the coupling fluid bottle on page 6 7 Waste bottle indicator A sensor monitors the level of waste fluid in its bottle Green OK The waste fluid bottle does not need emptying Red The waste bottle is full Empty the waste bottle see Empty and clean the waste water bottle on page 6 13 L Note Newer systems do not use the waste bottle Echo Liquid Handler Software 5 43 Focus calibration indicator Periodically the focal length of the acoustic transducer should be checked When this period
83. criptions illustrations and procedures apply to all liquid handlers unless the information is specifically stated for one model All models utilize ADE acoustic droplet ejection technology patented by Labcyte Inc Ultrasound based ADE precisely transfers nanoliter volumes of liquids including DMSO buffers with or without proteins or nucleic acids surfactants cell culture media and serum between microplates Excellent accuracy and precision in nanoliter volumes allow for direct transfer of compounds which reduces the need for intermediate plates and multiple dilutions and produces more accurate results in less Figure 1 1 time The Echo liquid handlers dispense as little as 2 5 nL without using pins tips or washing Echo 555 liquid handler Echo qualified source plates are specifically designed for the Echo liquid handler to take advantage of ADE technology and improve liquid transfer performance Nanoliter dispensing also makes an Echo liquid handler an excellent fit for many challenging life science applications in drug discovery genomics and proteomics Introduction 1 1 1 1 What is ADE Acoustic droplet ejection ADE technology is the process of transferring nanoliter volumes of liquid using acoustic energy The Echo liquid handler uses this technology to transfer fluid droplets from a source microplate to a receiving surface The following is a simplified sequence of ADE events 1 The Echo liquid handler
84. cs Calibration Advanced LABCYTE gt Protocol List 384 10nL 5 copies 10 0nl 384 384 384 to 1536 1 copy 2 5nl 384 153644 384 to 384 ts sy 5 0nl 384 384 Pr tocol Name 384 10nL 384PP Aqueous buff protein to 1536 Source Plate 384PP_AQ_SP f 384PP DMSO to 384PS 1 copy 500d Destination Plate 384PP_Dest Number of Copies 5 Transfer Volume 10 0nl 5 10 Echo Liquid Handler User Manual Echo software can be used to create several types of transfer protocols Examples of protocols that can be created with the Echo software are listed in the following table and described later in this section Table 5 1 Liquid transfer types Source Destination Transfer map type microplate microplate full or partial 384 well 96 well destacking quadrant destacking interleaved 4 regions from 1 src plate to 4 dest plates 2x2 or every other dest well 384 well 384 well standard 1 src plate to 1 dest plate 384 well 1536 well stacking quadrant stacking interleaved 4 src plates to 4 regions on 1 dest plate 2x2 or every other dest well stacking quadrant stacking interleaved ape wel 3456 well 9 src plates to 9 regions on 1 dest plate 3x3 or every third dest well destacking quadrant destacking interleaved 1536 well well 16 regions from 1 src plate to 16 dest plates 4x4 or every fourth dest well destacking quadrant destacking interleaved 4 regions from 1 src plate to 4 dest plates 2x2
85. d 4 Enter your laboratory information in the following screen Figure 2 3 Entering customer information f amp Echo 5XX Labcyte Inc x Customer Information Please enter your information User Name jsmith Organization labcyte Inc Install this application for Anyone who uses this computer all users Only for me jessie InstallShield 2 8 Echo Liquid Handler User Manual 5 Select the program location By default the Echo liquid handler software will install in the following folder C Program Files Labcyte Use the default location Figure 2 4 Selecting program location ie Echo 5XX Labcyte Inc Destination Folder Click Next to install to this folder or click Change to install to a different folder Install Echo 5XX to C Program Files Labcyte Change 6 Start the installation by clicking the INSTALL button Edit or verify the previously entered information or selections by clicking the BACK button Figure 2 5 Starting installation Ready to Install the Program The wizard is ready to begin installation Click Install to begin the installation If you want to review or change any of your installation settings dick Back Click Cancel to InstallShield Installation and Relocation 2 9 7 Close any running application Background processes like anti virus detection software can interfere with the installation Click the OK button to pr
86. d calibration For more information about manual operation read Chapter 4 Introduction 1 17 1 4 2 Modular workstation operation Modular workstation operation differs from manual operation with the addition of a robot to automate the loading and unloading of source and destination plates Typically a custom program is written that controls and synchronizes the Echo liquid handler and robot interactions An operator is still involved and performs the following functions Deliver master source microplates from the compound library Maintain a supply of empty destination microplates Select and execute the fluid transfer protocol Deliver the filled assay microplates to the next step in the process Monitor the status of the Echo liquid handler and the robot possibly with the assistance of a custom software program Figure 1 15 Modular workstation operation Echo Liquid Handler Workstation Master Empt Filled Source Destination _Assay Microplates Microplates Microplates 1 18 Echo Liquid Handler User Manual 1 43 Small cell operation Small cell operation includes the automation of processes other than liquid transfer Those processes might include several of the following m Lidding and delidding Microplate labeling Microplate sealing Incubation Reaction detection Temperature or humidity control Reagent addition Wash station used with other instruments Small cell mode operation Figure 1 1
87. d composition microplate properties and other factors A 4 Echo Liquid Handler User Manual Figure A 2 The ping BB TB and SR acoustic signals Ous 31 3us 32 0us 34 4 us Reflections Bottom of plate bottom Top of plate bottom Surface of well fluid From the survey signal measurements the following data is calculated m For DMSO water solutions the fluid composition is measured by interpreting the ratio of the amplitudes of the BB and TB echo signals BB stays nearly constant since neither the coupling fluid nor the microplate materials change impedance The TB echo on the other hand is directly affected by the DMSO water concentration A microplate well filled with 100 DMSO has a lower impedance than one filled with 70 DMSO water m The speed of sound through the well fluid is also directly affected by the fluid composition Once the concentration is known its speed of sound can be retrieved from a table in the Echo controller software m Fluid fill height can be measured once the well fluid s speed of sound and the echo return times are known The Echo liquid handler can measure the DMSO water concentration and calculate the well fill height for each individual source microplate well If the user knows the DMSO water concentration beforehand and knows that the well fill heights are consistent well to well quicker fluid transfer protocol options can be employed to improve throughput A source microplate survey can al
88. d on m The Echo liquid handler has been turned on but it has not completed its initialization m The Ethernet cable is not connected between the client PC and the Echo liquid handler m The Echo software is not configured to connect to the instrument IP address or hostname is missing or incorrect Check for the cause attempt to resolve the problem and try to launch the software again If the message persists call Labcyte Service and Support Echo Liquid Handler Software 5 3 5 2 Window description The Echo software starts with the first function the PROTOCOLS window This is the most frequently used function Figure 5 2 First window in the Echo software ABE 555 Liquid Handler localhost E3 Tabs for each p Protocols Labware Diagnostics Calibration Advanced function LABCYTE gt Protocol List Protocol list D a4 ion 5 copies 10 0ni 384 384 384PP Aqueous buff protein to 1536LDY 5 copies 2 5nl 384 1536 I 384PP DMSO to 384PS 10 copies 5 0nl 384 384 Function keys Add Edit Remove Cen 07 2240 14 23 57 807 SOAPFluidTransferProxy cpp 131 GetPlatelnfo START i 07 2240 14 23 57 823 SOAPFluidTransferProxy cpp 623 GetPlatelnfo END Server a n d cl lent 07 2240 14 23 57 823 SOAPFluidTransferProxy cpp 131 GetPlatelnfo START status window 57 SOAPFluidTransferProxy cpp 623 GetPlatelnfo END SOAPFluidTransferProxy cpp 131 GetAllProtocolHame SOAPFluidTransfer
89. ding the system or a system error DO Note Newer systems do not use the waste bottle 2 14 Echo Liquid Handler User Manual 6 Connect the chiller Attach the plastic tubing between the chiller and the Echo liquid handler Figure 2 1 Tubing connections on the chiller To chiller WATER IN From chiller WATER OUT A Caution Do not run the pump until the chiller tubing is connected Running the pump without the chiller tubing may damage other system components Figure 2 1 Tubing connections on the Echo system Fluidics Panel From chiller WATER IN To chiller WATER OUT Compressed air line AIR IN Vacuum Line VACUUM An 7 Connect the client PC with the Echo software to the Echo liquid handler with a cross over ethernet cable Fill the coupling fluid bottle and the fluidics circuit See Refill the coupling fluid bottle on page 6 7 and Cycle fluid through the Echo system on page 6 12 Installation and Relocation 2 15 2 16 Echo Liquid Handler User Manual CHAPTER SYSTEM DESCRIPTION This chapter covers the following Labcyte Echo liquid handler features and components m System overview e Echo front panel e Echo liquid handler back panel m Component description Process door Source plate gripper stage Source plate insert Destination plate gripper stage Status indicator lights LCD screen EMO switch Anti static bars AC power and fuse compartment
90. e Fluid leak indicator on page 5 42 Follow instructions on the LCD screen This message is generated by custom programming See your System Administrator 7 4 Echo Liquid Handler User Manual APPENDIX ADE TECHNOLOGY A 1 ADE history Simple ADE Acoustic Droplet Ejection liquid transfer using acoustic energy dates back to early experiments with high intensity acoustic beams at the Tuxedo Park laboratory of Alfred Lee Loomis in 1927 It was observed that immersing a high power acoustic generator in an oil bath would create a mound at the surface erupting oil droplets like a miniature volcano Improvements in the 1950s and early 1960s localized the energy with exponential acoustic horns or focused it with acoustic lenses but still maintained the high intensity of earlier devices These devices created drops with a continuous application of acoustic energy to form geysers of small droplets or patterns of disturbances on the liquid surface where some of the swells grew large enough to pinch off and become drops The introduction of a lower intensity process that both focused and pulsed the acoustic energy to create a single drop on demand was developed in the early 1970s Drop on demand technology has been extended to create today s ink jet printing The increasingly automated nature of life science research during the 1990s with its need for precise and reliable robotic liquid dispe
91. e If you cannot determine the original problem or you cannot correct the problem you must arrange for a Labcyte field engineer to restore your Echo liquid handler to normal use 4 6 Echo Liquid Handler User Manual 4 2 Define Labware Your Echo software was configured with Echo labware plate types prior to shipping If you need to add new or edit existing plates types select the Labware tab in the Main window and refer to the procedures in this section Figure 4 3 Labware window LA Echo 555 Liquid Handler localhost k Protocols Labware Diagnostics Calibration Advanced LABCYTE gt Plate Name List 1536HB Dest 1536HB_DMSO 1536LDV_Dest 1536LDV_DMSO 1536LDV_DMSO_HT Greiner _1536HiBasePS_782096 Greiner_384PS_781096 Add Edit Remove The source microplate types are specific to liquid types and use the naming convention platename_liquid type such as 384PP_DMSO Specialized features are appended to the plate name such as 1536_LDV_DMSO_HT This plate type is calibrated for high throughput HT dispensing which uses a faster transfer rate for higher volume or where precise droplet placement is not required See 1536 plate to 1536 plate liquid transfer on page 1 16 Since source microplates are carefully calibrated by Labcyte their plate types cannot be added or removed by the user and only the barcode location can be edited The destination microplate types are specific to liquid
92. e plate survey or liquid transfer accuracy will be affected 5 44 Echo Liquid Handler User Manual Maintenance mode Maintenance mode disables Echo motor control from the client software to allow you to perform maintenance procedures such as changing fluid Select the MAINTENANCE MODE checkbox to inhibit the execution of the following Echo processes and Maintenance Mode V functions from the Echo software Executing fluid transfer protocols Process door open close Source microplate extend retract Destination microplate extend retract flip Source microplate dryer up down on off Anti static bars up down on off Coupler nozzle up down Executing the dry plate diagnostic procedure Executing the move to well diagnostic procedure Executing the perform survey diagnostic procedure Homing the stages Power calibration Acoustic transducer calibration Barcode scanner calibration The following Echo functions continue to be available during maintenance mode m Creating editing or deleting fluid transfer protocol definitions m Creating editing or deleting destination plate definitions m Turning the coupler pump on or off or changing its pumping direction to normal or reverse m Executing either a hard or soft reset of the Echo controller Always clear the MAINTENANCE Mope checkbox when returning the Echo liquid handler to normal operation Echo Liquid Handler Software 5 45 5 7 3 Diagnostic procedures In
93. eers m Client Server Normal Mode installs the Echo client software onto the client PC and the Echo server software onto the Echo controller This software is installed by Labcyte field engineers Stub Mode creates a virtual Echo instrument onto the client PC that developers can use to develop and test custom software O Note Use this installation type only if you are writing your own programs that will call the Echo ActiveX programming interface Call Labcyte Service and Support if you have any questions regarding this installation type 1 Ethernet cable that switches or crosses over receive and transmit wire pairs that enable two de vices to communicate with each other 2 6 Echo Liquid Handler User Manual 2 3 1 Install the software The following procedure describes installation of Client Only software For information on installing Client Server software refer to the Labcyte Echo Integration Guide To install Echo liquid handler software 1 Insert the Echo liquid handler software installation CD into the CD drive The installation should automatically start If the installation does not start a Go to My Computer and double click the drive icon for the CD drive b Look for setup exe in the directory on the CD and double click the icon 2 Select Install Client Only and enter the Host Name or IP address that will be used by the Echo liquid handler The Echo liquid handler comes pre configured with IP address 1
94. efinition editing is not restricted Remove To remove an existing destination plate definition select the definition name and click the REMOVE button The selected destination plate definition will be deleted 5 30 Echo Liquid Handler User Manual Manage labware definitions Adding a new destination plate definition or editing an existing definition are virtually the same process On the LABWARE tab click either ADD or Epit The DESTINATION PLATE SPECIFICATION dialog box will open If an existing definition is being edited the PLATE NAME will not be editable O Note The edit window for the source plate specification looks the same but only the barcode location can be edited Figure 5 1 Plate Specification for Destination Plate Plate Name itis Manufacturer pooo LotNumber PartNumber Rows 16 Columns 24 Barcode Location None A1x Offset A 1 mm aA1vOffset 1 mm X Center Spacing C 45 mm Y Center Spacing D 45 mm Plate Height E 14 35 mm Flange Height F 2 41 gt mm Well width G 0 01 mm Well Capacity H 0 01 u Plate Name Enter a plate name to identify the new definition required Manufacturer Enter the manufacturer s name Lot Number Enter the plate lot number identification Part Number Enter the manufacturer s plate part number Rows Enter the number of rows The valid range is one or more Columns Enter the number of columns The valid range is one or mo
95. er which involves a human operator performing the following procedures m Startup Shutdown m Define Labware m Create liquid transfer protocol m Start liquid transfer run This chapter takes the reader through the basic steps necessary to transfer a compound from one microplate to another with the Echo liquid handler It is intended to present the entire range of activity in a start to finish flow It is not intended to provide all of the information necessary to understand each individual step Refer to the chapter Echo Liquid Handler Software on page 5 1 for a detailed description of each software function Manual Operation 4 1 41 Startup Shutdown At this point the hardware and client PC software installations should be complete All fluid air and electrical connections should be complete and functioning Your Labcyte field engineer performs a set of function tests to ensure that the system is operating correctly 41 1 Turn on Echo power Turn on the Echo liquid handler by pressing the AC power switch located on the back panel The following message may appear while the Echo liquid handler initializes Figure 4 1 Connection failure message Failed To Connect To Echo Server e x i Host refused connection Verify address and port number localhost Click OK to try to connect to server again click Cancel to exit Echo Liquid Handler x IE Possible causes for connection failure include
96. er ECHO AL 15 R 50 uL of algaecide Labcyte catalog number ECHO AL04 Gloves for protection when handling the algaecide Paper towels or shop cloths to deal with any unexpected fluid leaks Screwdriver for removing the fuse cover The fluidic system will be emptied purged and refilled several times therefore take note of the following caution A Caution When working with the Echo coupling fluid system and turning its coupling fluid pump on and off make sure that the vacuum line is connected and working before turning the coupling fluid pump back on If the vacuum line is disconnected or the vacuum system is not operating correctly when the coupling fluid pump is turned on the coupling fluid can flood the bottom of the system Maintenance and Service 6 9 6 2 4 Empty the coupling fluid bottle 1 Ensure that no fluid transfer or diagnostic procedures are under way 2 Turn off the vacuum and coupling fluid pump a Disconnect the vacuum line from the Fluidics Panel to decrease vacuum pressure b Turn off the coupling fluid pump through the Diagnostics windows of the Echo software O Note The surge tank will add a few seconds to stopping the vacuum 3 Unscrew the cap from the coupling fluid bottle Figure 6 1 Remove m Sree Vacuum line connected to fluidics panel 4p TEE Coupling fluid bottle 4 Remove the coupling fluid bottle from its caddy and empty the bottle 5 Replace the cap and its attached tubin
97. es in the section Instrument Relocation on page 2 12 A Caution Do not store the Echo liquid handler outside of the storage temperature range 5 C 45 C 41 F 113 F Temperatures outside of this range can damage the instrument Maintenance and Service 6 19 6 20 Echo Liquid Handler User Manual CHAPTER CONTACT INFORMATION AND TROUBLESHOOTING This section includes setup troubleshooting and maintenance alert information 7 1 Contact information If you need help in the following areas m Technical support m Applications support m Service m Repair m Parts supplies and accessories Please contact Labcyte Technical support line 408 747 2000 option 8 or 408 542 7866 Email support Labcyte com Address Labcyte Inc 1190 Borregas Ave Sunnyvale California 94089 877 742 6548 408 747 2000 Contact Information and Troubleshooting 1 1 7 2 Setup problems If the Echo software cannot connect with the Echo liquid handler the following message will appear Figure 7 1 Connection failure message Failed To Connect To Echo Server 14 A x i Host refused connection Verify address and port number localhost Click OK to try to connect to server again click Cancel to exit Echo Liquid Handler This message is commonly seen and is not a cause for alarm Possible causes for the connection failure include m The Echo liquid handler has not been turned on m The Echo liquid handler
98. eturning to the coupling fluid bottle and that it does not contain a large number of bubbles 2 To completely purge the Echo fluidic circuit repeat the empty purge fill procedure two more times 3 After filling the fluidic circuit the coupling fluid bottle will be partially empty Turn the coupling fluid pump off and add coupling fluid so that the bottle contains 875 mL of coupling fluid Make sure that the J shaped tube isn t covered by the new fluid level 4 If repeated empty fill cycles have been performed remember to add algaecide at the ratio of 50 uL per 1000 mL of distilled water 5 Turn on the coupling fluid pump 6 Check the COUPLING FLUID status on the Echo software DIAGNOSTICS window The indicator should be green E Note If the empty purge fill procedure is being performed more than once add the algaecide only during the last fill of the coupling fluid bottle 6 12 Echo Liquid Handler User Manual Empty and clean the waste water bottle The Echo liquid handler dries each source plate before it is extended outside of the process door To dry the source plate the Echo liquid handler vacuums coupling fluid from the bottom of the source plate and sends the fluid to the waste water bottle DO Note Newer Echo systems do not use a waste water bottle Empty the waste water bottle when any of the following occur m The WASTE BOTTLE indicator on the DIAGNoSTICS tab of the Echo software is red indicating that the waste
99. f automated system integrated configurations This section explores the following operating modes Manual mode Modular workstation mode Small cell mode Fully automated mode The Echo server client architecture allows multiple client computers programs to connect to an Echo liquid handler at the same time Read Multi client considerations on page 1 21 for a discussion of the advantages and disadvantages of multiple client operation E Note The Echo software comes with a static IP address 192 168 1 1 If this IP address is not available or if you are installing more than one Echo liquid handler on a network contact your Labcyte field engineer to assign different IP addresses to your Echo devices a separate IP address is required whenever more than one Echo liquid handler is installed 1 4 1 Manual operation Manual operation leaves control of every step to the operator m Defining new fluid transfer protocols or new destination microplates m Executing a fluid transfer protocol m Loading and unloading both source and destination microplates m Monitoring Echo status Figure 1 14 Manual operation Master Empt Reformatted Client Source Destination Echo _Assay Workstation Microplates Microplates Liquid Handler Microplates Manual operation is also used when performing the following functions m Defining new destination microplates m Defining new fluid transfer protocols m Performing periodic maintenance an
100. g basin uid inlet transducer y RF Generator Coupling fluid outlet A 4 4 Examples of acoustic droplet ejection Figure A 5 shows a 5 nL droplet being ejected from a volume of water The acoustic energy pulse creates a mound of fluid at the surface which is pushed up A droplet forms on a neck and eventually pinches off Its momentum carries it up to a receiving surface while the fluid mound recedes into the larger fluid volume Figure A 5 A droplet ejection A 8 Echo Liquid Handler User Manual Each image in Figure A 6 shows a stroboscopic sequence of a 5 nL DMSO droplet ejection Each 5 nL 212 um diameter droplet is traveling upward at approximately 1 m s to a glass slide In each of the four images the droplet has been photographically captured at six points in time by a multi flash strobe at intervals of 400 us The four images show 1 20 40 and 100 drops deposited on the slide Figure A 6 Multi stroboscopic image view of ADE et et es ws A 9 A 10 Echo Liquid Handler User Manual APPENDIX BARCODE LOCATIONS The Labcyte Echo liquid handler optionally includes one or more barcode scanners The location of each scanner depends on the location of your barcode labels on the microplates CS Note You can use either the front scanner or the side scanner but not both The load registration pins on the source and destination microplate assemblies obstruct the barcode scanners from seeing all of a microplate s front a
101. g onto the empty coupling fluid bottle Return the coupling fluid bottle to its caddy Purge the fluid circuit 1 Turn off the coupling fluid pump 2 In the Diagnostic window of the Echo software select the reverse coupling fluid pump direction COUPLING PUMP DIRECTION REVERSE See Coupler pump direction normal reverse on page 5 40 3 Turn on the coupling fluid pump via the pump power switch The coupling fluid inside the Echo fluidic circuit will be pumped back into the coupling fluid bottle Bs Note The fluid returning to the bottle will raise the water level therefore watch for potential overflow in the coupling fluid bottle 4 Turn off the coupling fluid pump 5 Return the coupling fluid pump direction to normal Select COUPLING PUMP DIRECTION Normal 6 10 Echo Liquid Handler User Manual 6 Unscrew the cap on the coupling fluid bottle and remove the bottle from the bottle caddy Clean and refill the coupling fluid bottle 1 Clean the coupling fluid bottle and tubes a Add Alconox or hand soap and water to the bottle b Scrub the bottle and tubes with a plastic bristle bottle brush c Thoroughly rinse the bottle and tubes to ensure there is no soapy residue Use distilled water for the final rinse d Allow the bottle to dry if possible or alternate with another 1000 mL bottle Note If you see foam while refilling the bottle rinse the bottle and tubes again with hot tap water 2 Inspect the
102. ge 5 13 Manual Operation 4 11 4 4 Run the liquid transfer protocol Before running the liquid transfer protocol you will need to prepare the compound to be transferred and set up the microplates 4 4 1 Prepare the microplates Prepare the microplates as follows 1 Prepare the source microplates Consider the following factors m 100 DMSO absorbs water rapidly from the atmosphere and can increase the final volume of the compound in the source microplate which might exceed the maximum volume allowed by the Echo liquid handler m Aqueous solutions evaporate quickly at low volumes For longer runs fill wells to the maximum volume specified for the source plate type used Volume ranges may differ for different aqueous solutions as well as aqueous vs DMSO based solutions m Ifthe source microplates were made earlier and stored refrigerated or frozen allow them to equilibrate or thaw to room temperature 2 Place the appropriate plate insert onto the plate gripper if it is not already in position See Source plate insert on page 3 6 3 Collect the microplates that will serve as destination plates to receive the compound A destination microplate definition must exist or be created For example knowing the plate height of the destination microplate allows the Echo liquid handler to lower the destination microplate into very close proximity to the source microplate improving droplet ejection accuracy and reliability
103. has been turned on but it has not completed its initialization e Check the gripper stage homing sequences particularly the theta rotation of the destination gripper stage e Check the door opening sequence m The Ethernet cable is not connected between the computer and the Echo liquid handler m The Echo software is not configured IP address or hostname to connect to the instrument m The Echo liquid handler is failing to communicate Check for the cause attempt to resolve the problem and try to launch the software again If the message persists call Labcyte Service and Support 7 2 Echo Liquid Handler User Manual Maintenance alerts During normal use the Echo liquid handler may prompt that it needs attention Refer to the following maintenance alerts Table 7 1 Echo Maintenance Alerts Maintenance Alert Procedure Q Warning light is yellow Possible causes See Status indicator lights on page 3 8 Q The Echo stages acoustic transducer source plate and destination plate are not homed Q The fluid level is low in the coupling fluid bottle Q The coupling fluid temperature is outside of the normal range 21 2 C 23 0 C Perform Home calibration See Home calibration on page 5 52 Add more fluid See Refill the coupling fluid bottle on page 6 7 Check the chiller setting it should be at 22 C See Coupling fluid temperature and indicator on page 5 44
104. ibrated for 1536 well source plates you can create a new 1536 1536 protocol definition using the ADD NEW PROTOCOL WIZARD as described above for 384 384 protocols with the following exceptions m An existing 1536 well source microplate must be selected m The transfer volume range may be different than those used with 384 well source plates For example for the Echo qualified 1536 well source microplate the valid range is 2 5 to 2000 nL in increments of 2 5 nL If you type an invalid transfer volume you will receive an error message that displays the correct transfer range Figure 5 6 Invalid transfer volume for 1536 well plate x The valid range For transfer volume is from 2 5 to 2000 0 nL OK Create a new 384 1536 protocol definition Create a new 384 1536 protocol definition using the stacking features of the ADD New PROTOCOL WIZARD Compound from four 384 well source plates can be transferred to a single 1536 well destination plate See 384 plate to 1536 plate liquid transfer on page 1 14 for an illustration of plate mapping to more than one destination plates L Note ifthe new liquid transfer protocol requires a new destination microplate definition create it before running the ADD NEW PROTOCOL WIZARD See Manage labware definitions on page 5 31 1 Click the ADD button to start the ADD NEW PROTOCOL WIZARD Enter the following information Protocol Name Enter a name for the new protocol It must not a
105. ical interference or misalignment When a problem is detected the following actions occur e The coupling fluid pump immediately turns off depending on the problem e Front and rear panel fault lamps and LCD messages notify the operator that a problem has occurred e ActiveX events notify remote systems that are connected to the Echo liquid handler 1 6 Echo Liquid Handler User Manual 1 2 3 Ease of use An Echo liquid handler is quickly ready for use m Minimal operator training is required m Preparation for liquid transfers requires little more than defining a transfer protocol and selecting the source and destination plates being used Protocols or destination plates can be quickly defined by the operator m Liquid transfer requires only a few clicks by the operator using the Echo software which is included with each instrument m The Echo liquid handler can be operated manually by a researcher or as part of a fully automated drug discovery lab For example Figure 1 5 demonstrates the simple two step process required to run an existing fluid transfer protocol using the Echo software program Figure 1 5 Executing a transfer protocol in two steps LABCYTE gt Protocol List 384 10nL 5 copies 10 0nl 384 384 384PP Aqueous bufF protein to 1536LDV 5 copies 2 5nl 384 1536 I 384PP DMSO to 384P5 10 copies 5 0nl 384 384 TS Select a protocol Run the protocol Add Edit Rem
106. ices interconnected on an Ethernet LAN local area network Please review the following information and take appropriate measures to avoid conflicts caused by multiple workstations controlling an Echo liquid handler inappropriately The Echo liquid handler can be connected to a network or to more than one workstation This configuration allows one or more users to control the Echo remotely to download liquid transfer information directly to the network or to connect more than one Echo liquid handler to an automated filling operation Figure 1 18 Multiple clients talking to an Echo liquid handler Ethernet Operator s _ Echo Supervisor s Workstation Liquid Handler Workstation For example a local workstation might be used to do the following m Monitor Echo instrument status m Perform user maintenance m Execute troubleshooting procedures At the same time a central system may be used to control all of the day to day work flow involving the instruments in the lab In relation to an Echo liquid handler it might be involved in the following m Define fluid transfer protocols m Define destination microplates m Deliver source and destination microplates to the Echo liquid handler via conveyor belt and robot m Remove source and destination microplates m Execute fluid transfer protocols m Handle all of the data The Echo liquid handler is a client server system The client is the external workstation that contains the
107. id transfer using the Echo software the first step is to create a protocol A protocol defines the source and destination microplates to use the volume to transfer number of transfers and the specific wells involved Protocols are stored in the Echo database and displayed every time you view the PROTOCOLS window From the PROTOCOLS window you can click the ADD button to begin the Add New Protocol wizard After you have created the protocol you can select it in the Protocol List see Figure 5 1 to run edit or remove Figure 5 1 The protocol list LABCYTE gt Protocol List 384 10nL 5 copies 10 0nl 384 384 384PP Aqueous buff protein to 1536LD 5 copies 2 5nl 384 1536 I 384PP DMSO to 384PS 10 copies 5 0nl 384 384 Add Edit Remove Samples of protocols are described in the sections that follow 5 12 Echo Liquid Handler User Manual Create a new 384 384 protocol definition Create a new 384 384 protocol definition using the ADD NEw PROTOCOL WIZARD O Note If the new liquid transfer protocol requires a new destination microplate definition create it before running the ADD NEw PROTOCOL WIZARD See Manage labware definitions on page 5 31 1 Click the ADD button to start the ADD NEW PROTOCOL WIZARD Enter the following information Protocol Name Enter a name for the new protocol It must not already be in use m Source Plate Type Select a source plate type from the first drop down list
108. ill height in a microplate well is measured by the Echo liquid handler and used to focus the acoustic energy near the well fluid s meniscus Changes in fluid composition over time Evaporation can cause significant changes in well volume especially in warm or dry environments In addition concentration in a DMSO based solution changes over time due to hydration If the DMSO water ratio is known the transfer protocol definition can include a concentration map When the DMSO water ratio is unknown it must be measured by the Echo liquid handler The Echo liquid handler uses dynamic real time measurement of the fluid properties at multiple times during a run to respond instantly and automatically to changes in the wells It measures the fluid and surface properties During the survey at the beginning of the run Immediately before each well transfer Immediately after each well transfer A 2 Echo Liquid Handler User Manual A 3 Source microplate survey Prior to each run the Echo liquid handler scans the fluid in each source microplate well During the microplate survey the acoustic transducer is moved beneath each well A pulse of acoustic energy is transmitted up through the bottom of the microplate into the fluid in the well The low energy acoustic pulses used during the source plate survey will not eject droplets of fluid The three echo signals When an acoustic pulse commonly referred to as a ping is transmitted the
109. in a minimum operating envelope as follows see Figure 2 1 Operating space envelope 59 cm 23 2 in width x 99 cm 39 in depth x 118 cm 46 5 in height Figure 2 1 Instrument layout on bench top T Top clearance 25 4 cm 10 in Operating Envelope 118 cm 46 5 in Side clearance Za Rear clearance 30 5 cm 12 in Sos a Br a K 2 oF gt n Pa Front clearance SX do Side clearance es Pa 7 6 cm 3 in RS p 2 5 cm 1 in Top clearance 25 4 cm 10 in minimum above the instrument to remove the top cover for servicing CS Note Less height clearance gt 2 5 in is acceptable if there is sufficient space to lift the top and slide it all the way forward Talk to your Labcyte field engineer Front clearance sufficient clearance to load and unload microplates by hand or by robot Side clearance 2 5 cm 1 in minimum on left and right sides 2 2 Echo Liquid Handler User Manual Rear clearance 30 5 12 in minimum behind the instrument to allow for cables hoses tubing and maintenance access Note The coupling fluid bottle and caddy fit within the rear clearance If you position the bottle and caddy on the side of the instrument add at least 17 8 cm 7 in to the side clearance Additional space for the client PC coupling fluid chiller if it is on the bench and microplates Additional space would also be needed if you use robotics AC power The user must provide AC power mat
110. in the inner left corner Click the NEXT button Run Protocol 384 10 nL 1 copy 10 0nl 384 384 Please load source plate aggregate 1 of 1 Pressing Next will retract the source plate and present the destination gripper The source plate gripper stage retracts into the Echo liquid handler and the destination plate gripper stage is extended outside of the Echo liquid handler 4 Load your destination microplate with well A1 in the inner left corner Click the Next button Run Protocol 384 10 nL 1 copy 10 0nl 384 384 Please load destination plate 1 of 1 Pressing Next will retract the destination gripper and the transfer will begin While the destination plate gripper stage retracts the microplate the source microplate is surveyed The Echo liquid handler determines the fluid composition and well fluid thickness and then calculates the fluid volume in each well 5 26 Echo Liquid Handler User Manual When the survey is complete and the destination microplate is positioned correctly above the source microplate the liquid transfer process begins Using the data contained in your destination microplate definition the protocol definition and the source microplate survey just performed droplets are ejected from each specified source microplate well into the corresponding destination microplate well A progress bar keeps you informed of the liquid transfer in progress You can abort the liquid transfer process by
111. inations To select an entire row or column click the appropriate header To select all wells click corner header or press ctrl A C Dest 1 B C Dest2 C D E C Det3 F G H C Dest4 I 1 K all L M H 2 l The DEFINE WELLS dialog box shows a simple grid that represents the microplate Each source microplate well is represented by a grid square The user selects the microplate wells from which fluid will be transferred 5 22 Echo Liquid Handler User Manual Selected wells highlighted in blue will be transferred when the protocol is run The dialog box shown above defines a liquid transfer from wells A1 through 16 There are several methods to define which wells belong in the transfer map Click a single well to select it and clear all other selected grid squares Hold the CONTROL key down and click in individual wells Each well is selected in turn After making a well selection hold down the SHirr key and click in another well All wells in a rectangle defined by the two wells are selected Click on a row label or a column label The entire row or column will be selected Click and drag the mouse point to define a rectangle of wells in one move If a group of wells is selected and another well or group of wells must also be selected hold the CONTROL key down so that the original group of wells is not cleared All wells can be selected by clicking in the upper left square above the row labels Another
112. ine hose barb for 9 5 mm 3 8 in ID tubing included in Echo liquid handler accessory kit Chiller performance 22 1 C 0 9 C 71 8 F 1 6 F 3 3 3 Environmental Temperature operating 21 C 5 C 70 F 9 F Temperature storage 5 C 45 C 41 F 113 F Humidity operating 10 80 non condensing Mounting surface Sturdy stable capable of handling 128 kg 283 lbs weight Pollution degree 2 Installation category 3 3 4 Electrical There are two electrical configurations of the Echo liquid handler Config Voltage Frequency Power Fuse A 100 120 VAC 50 60 Hz 10A 250 VAC 10 A Fast Blow B 200 240 VAC 50 60 Hz 5A 250 VAC 5A Fast Blow 3 3 5 Client PC Operating system Microsoft Windows XP SP2 or later SOAP SDK ver 3 0 or later CPU Intel Pentium 4 or later Memory 256 MB or higher Hard drive storage 200 MB Video 1024 X 768 resolution 256 colors or better Network connection 10 100BaseT Network protocol TCP IP User account Labcyte API ActiveX Programming languages C C C Visual Basic Visual J or NET compatible 1The client PC must have a Labcyte account with administrator privileges that is accessible by LabcyteService and Support to service the Echo liquid handler system System Description 3 19 3 3 6 Communications Network connection 10 100BaseT Network protocol TCP IP 3 3 7 Fluid transfer Volumetric Accuracy lt 10 error single so
113. ism movement is useful during maintenance troubleshooting and diagnostic procedures Access to device control is found in several areas of the Echo software m The Device Control and Status section of the DIAGNOSTICS window uses eleven sets of checkboxes and status indicators They are discussed below m The Miscellaneous section of the DIAGNOSTICS window uses the diagnostic procedures drop down list and LAUNCH button See Diagnostic procedures on page 5 46 m The MOTION section of the CALIBRATION window uses the HOME and REGISTER buttons See Motion calibration on page 5 52 L Note The Echo liquid handler will not allow device movement that would cause damage to itself For example if the process door is closed when the source plate gripper stage is commanded to extend outside the Echo liquid handler opens the process door first Figure 5 1 Device control and status section m Device control and status Door V Open M Close Q Source Plate Vin FM our D Present Destination Plate V In M our Q Present Flip Destination Plate Up Q MN Down Plate Dryer Up O V Down Plate Dryer F On O vor Anti static Bars on O Vor Upper Anti static Bar JV Up I Down Q Coupler Noze Up Q V Down Coupler Pump M on M off QO Coupler Pump Direction IV Normal D Reverse 3 Peters EE ee Door open close The Echo door can be manually opened and closed by selecting the appropriate checkbox If either the s
114. l A vlolzlziclzx ls zlaln mlolo al gt 4 Number of Copies Enter the number of times the protocol will be executed The valid range is 1 to 10 000 copies The operator is prompted to load and unload the source microplate and the specified number of destination microplates 5 14 Echo Liquid Handler User Manual 5 Transfer Volume Enter the total volume of fluid to be transferred The valid range is dependent on the source microplate see Maximum Transferred Volume on page 3 21 For example for the Labcyte 384 well PP source microplate the valid range is 2 5 to 10000 nL in increments of 2 5 nL2 If you enter the transfer volume directly instead of using the increment decrement buttons remember that the value must be an even multiple of the minimum transfer volume If you type an invalid transfer volume you will receive an error message that displays the correct transfer range Figure 5 4 Invalid transfer volume for 384 well plate x The valid range for transfer volume is From 2 5 to 10000 0 nL OK 6 Click the FINISH button to complete the new protocol Figure 5 5 384 384 protocol finished Add New Protocol Wizard Page 2 of 2 Transfer Wells A1 16 View Define Number of Copies 1 E Transfer Volume 10 nL m 2 The valid range may also differ for different fluid types Echo Liquid Handler Software 5 15 Create a new 1536 1536 protocol definition If your Echo liquid handler has been cal
115. l Enabling power monitoring adds significant overhead to each droplet ejection and decreases liquid transfer throughput Default Power monitoring off Save Plate Survey Data to Disk The data collected during a source plate survey is held in ee a cache memory It is replaced when the next source plate survey is performed If the SAVE PLATE SURVEY Data TO Disk checkbox is selected the survey data will be stored on the client PC disk The data file is stored to lt InstallDir gt Labcyte Medman Data This data can be useful to Labcyte personnel during diagnostic troubleshooting Default Source plate survey data is not saved on the Echo controller 5 64 Echo Liquid Handler User Manual Save Transfer data to disk Normally the data collected during a source plate transfer is deleted when the transfer is complete If the SAVE TRANSFER DATA TO Disk checkbox is selected the transfer data will be stored on the client PC disk There is no reason to save transfer data during normal operation The data file is stored to lt InstallDir gt Labcyte Medman Data Transfer data can be useful to Labcyte personnel during diagnostic troubleshooting Default liquid transfer data is not saved to the Echo controller Ignore Source Plate Sensor Check Selecting the IGNORE SOURCE PLATE SENSOR CHECK checkbox bypasses the source plate present sensor This may be useful when an individual source plate does not fit correctly A run protocol procedu
116. l locations applies to all models XIV Echo Liquid Handler User Manual Product label The Echo liquid handler product label includes Labcyte corporate identification product identification product serial number regulatory agency marks origin of manufacture date of manufacture input power specifications and CE mark J LABCYTE gt Sunar CA 94089 Serial Date of Manufacture Manufactured in the United States Power Rating Fuse Rating This product conforms to the applicable requirements of 21CFR subchapter J at the date of manufacture ETL label or The ETL label indicates conformance to ETL ETL LISTED SEMKO product safety standards for the US CONFORMS TO id anaes eN aA US LST G1010A 1 3084885 CERTIFIEU TO CAN CSA STD C22 2 NO 1010 1 Intellectual property label The intellectual property label cites United States patents and other applicable legal protection Electrical hazard The electrical hazard label warns of possible injury from electrical cables connections and circuits inside the Echo liquid handler 3 The CE mark indicates compliance with the EU European Union Directives Preface XV xvi General warning label The general warning label reminds users to take normal safety precautions when operating and working around an Echo liquid handler Laser safety label The laser safety label warns that the Echo liquid handler optionally uses laser based barc
117. l of the default values and do not select the OVERRIDE CALCULATED RANGE checkbox 3 Click the NEXT button The Echo process door opens the source plate gripper stage extends and the Echo liquid handler performs a focal sweep The focal sweep uses acoustic pings bounced off an internal registration plate while the transducer is being moved vertically The focal sweep process measures the transducer s focal length After the focal sweep is done the bottom of the registration plate is dried 4 Click the FINISH button to complete the focus calibration procedure Figure 5 2 Focus calibration page 2 new focal length calibration not required Position vs Amplitude Results 3 55195 Start ToF us 33 08 Stop ToF us 34 42 3 42492 Previous Focus ToF us 33 756 New Focus ToF us 33 774 E 329789 Previous New us 0 0176 3 E a 3 17086 3 04383 2 9168 31 4336 32 3579 33 2821 34 2064 35 1306 36 0549 Press Finish to save the new Focus ToF ToF us Press Back to retract source gripper lt Back Cancel 5 58 Echo Liquid Handler User Manual Note If your graph shows two curves the Echo software has detected older calibrations software version 2 2 and older and has performed a focus calibration check for both current and older calibrations Position vs Amplitude Results 347813 Vi v2 Start ToF us 32 92 32 92 3 30908 Stop ToF us 34 27 34 27 Previous Focus ToF us 33 600 33 6006
118. lect Survey from the MISCELLANEOUS drop down list Maintenance Mode _ 3 Click the LAUNCH button The SURVEY window will open Figure 5 2 Survey procedure Survey Eile Volume uL Results Fluid DMSO 70 100 Percent Volume Start Well Al Fluid Composition Number of Rows 16 Fluid Thickness Number of Columns 24 Rav Ray Save Results to Disk M Processed homogeneaus G Processed inhomogeneous Ignore Source Plate Sensor 4 From the NAME drop down list select the correct source plate type Echo Liquid Handler Software 5 47 5 Enter the wells to be surveyed Use one of the following methods m Enter the values in the Parameters section a Enter a valid starting well Example A1 b Enter the number of rows to be surveyed Example 16 c Enter the number of columns to be surveyed Example 24 m Use the mouse to select the wells in the plate map 6 Set up the following options in the PARAMETERS section as appropriate Save Results to Disk Select this checkbox to automatically store the results of the acoustic survey in the client PC data directory C Program Files Labcyte Medman Data Also see FILE SAVE on page 5 50 Ignore Source Plate Sensor Select this checkbox to resolve problems with fitting a source plate onto the source plate gripper stage This option allows the Echo liquid handler to use a source plate without detecting it
119. ler directly to the client PC or to a network This connection allows you to monitor and control the Echo liquid handler If the Echo liquid handler is connected directly to a client PC the Ethernet cable must be a cross over cable Other data connection ports are used by Labcyte field engineers Figure 3 2 External connections power and data MOR ETHERNET w RJ 45 Ethernet port 3 14 Echo Liquid Handler User Manual 3 2 10 Coupling fluid bottle The Echo system comes with a 1000 mL coupling fluid bottle See Figure 3 1 CS Note Older Echo systems are also equipped with a 500 mL waste bottle Figure 3 1 Coupling fluid bottle Next to the coupling fluid bottle is a sensor that is connected by cable to the Echo liquid handler The sensor monitors fluid level When the fluid falls below a specific level the sensor sends a low fluid volume alert to the status panel See Figure 3 2 Figure 3 2 Coupling fluid bottle and sensor connections Coupling Fluid Sensor Coupling Fluid Bottle System Description 3 15 3 2 11 Fluid chiller Another essential component of the Echo liquid handler system is the fluid chiller This component draws water from the coupling fluid bottle adjusts the fluid temperature and circulates the fluid through the acoustic transducer Coupling fluid temperature control Correct Echo liquid handler coupling fluid temperature is 22 0 C This temperature can be monitored o
120. lities are being added regularly Check with your sales representative for more information Introduction 1 11 iquid transfer Figure 1 8 and Figure 1 9 below illustrate simple fluid transfers between two 384 well microplates While Figure 1 8 shows a full a multi region plate to 384 plate 384 1 3 1 plate transfer Figure 1 9 shows partial plate transfer Both transfer protocols are easily defined by the 1 operator using the Echo software program Figure 1 8 384 to 384 full plate transfer Figure 1 9 384 to 384 multi region partial plate transfer Figure 1 10 illustrates a more complex fluid transfer between two 384 well microplates Droplets are ejected from multiple source plate wells into the same destination plate well This transfer map and its transfer protocol cannot be defined but it can be defined with a custom program and Echo ActiveX programming interface refer to Labcyte Echo Integration Guide 1 using the current Echo software program Figure 1 10 384 to 384 multiple source plate wells into one destination plate well transfer 1 12 Echo Liquid Handler User Manual 1 3 2 384 plate to 96 plate liquid transfer Figure 1 11 illustrates a 384 well to 96 well quadrant fluid transfer Wells from a single 384 well plate can be transferred or destacked to four 96 well plates Individual wells or all wells from each quadrant can be transferred to the corresp
121. ll motors are disabled The coupling fluid pump is turned off All pneumatic actuators are sent to their default positions The anti static bars are turned off All RF devices RF power supply RF amplifier RF switch and so on are turned off m The HV grid is turned off m The internal controller aborts all processes in progress returns an error to the system invoking the aborted process designates the three stages as not calibrated and goes into a halted state Activating the EMO switch does not turn off the following Internal Echo liquid handler controller AC power inside the Echo liquid handler power distribution panel Internal 24 VDC power supply power output however is interrupted External communications Ethernet USB Coupling fluid chiller Manual Operation 4 5 Recovering from an EMO shutdown If the EMO switch has been used to shutdown the Echo liquid handler use the following procedure to start up again 1 Determine and resolve the original problem 2 Turn the Echo liquid handler back on by twisting the EMO switch clockwise It will pop out to its normal operating position When the EMO switch is returned to its normal position power is again supplied to most internal systems and most internal mechanisms are enabled The Echo liquid handler stage motors however are not enabled yet the operator must explicitly command a stage homing procedure see Motion calibration on page 5 52 CS Not
122. lready be in use m Source Plate Type Select a source plate type from the first drop down list m Destination Plate Type Select a destination plate type from the second drop down list The destination plate definition must already exist in the Echo database 5 16 Echo Liquid Handler User Manual Figure 5 1 Add new protocol wizard page 1 of 2 Add New Protocol Wizard Protocol Name 384 to 1536 Source Plate Type 384PP_DMSO z Destination Plate Type z 2 Click the NEXT button to open the second ADD New PROTOCOL WizARD dialog box Figure 5 2 Add new protocol wizard page 2 of 3 Add New Protocol Wizard Page 2 of 3 q r Transfer Map Type Quadrant Transfer Volume 2 5 nl E Enter the following information m Transfer Map Type Select an interleaved or a quadrant liquid transfer protocol type See What types of liquid transfer are supported on page 1 11 Number of Copies Enter the number of times the protocol will be executed The valid range is 1 to 10 000 copies You are prompted to load and unload the source microplate and the specified number of destination microplates m Transfer Volume Enter the total volume of fluid to be transferred The valid range is dependent on the source microplate see Maximum Transferred Volume on page 3 21 For example for the Labcyte 384 well PP source microplate the valid range is 2 5 to 10000 nL in increments of 2 5 nL If you enter the tran
123. m Inspect and clean the grid on the destination plate gripper See Clean the destination plate gripper grid on page 6 16 m Inspect the coupling fluid tubing for algae growth If growth is present replace the tubing and purge the system Some yellow discoloration is acceptable however heavy discoloration indicates a problem Replace the tubing and purge the system See Purge the fluid circuit on page 6 10 Annual preventive maintenance Labcyte will provide annual preventive maintenance Please talk to your Labcyte service representative or call Labcyte Service and Support to schedule the date If you are no longer in warranty or have a service contract contact Labcyte to obtain a quote and schedule preventive maintenance See Contact information on page 7 1 Maintenance and Service 6 5 6 2 Maintenance procedures A Warning Do not attempt to service or repair the internal Echo mechanisms yourself The electrical laser and mechanical systems must be maintained by trained personnel Other than the user level maintenance procedures described in this chapter all service and repair must be performed by Labcyte field engineers If you have questions regarding what may be serviced by an Echo user please call Labcyte Service and Support See Contact information on page 7 1 6 2 1 Maintenance tools and materials Tools required The following tools or items are required during Echo maintenance procedures
124. n the DiAGnostics tab of the Echo liquid handler software application See Coupling fluid temperature and indicator on page 5 44 Set the fluid chiller to 22 0 C See Appendix C for chiller instructions A Caution Incorrect fluid temperature can reduce the precision and accuracy of the fluid that is transferred and potentially damage the acoustic transducer Ensure that the chiller is correctly connected to the Echo instrument and is running at recommended temperature 3 2 12 Surge tank A 10 L surge tank is available upon request When vacuum demand is at its highest the surge tank ensures a continuous source of vacuum It is especially useful in laboratories where house vacuum barely meets the minimum vacuum requirement or is inconsistent due to other laboratory demands 3 16 Echo Liquid Handler User Manual 3 2 13 Fluidics panel The fluidics panel contains tubing and hose connections to the following components Figure 3 1 Fluidics panel WATER IN 24 WATER OUT s To chiller se gt SF AIR IN From chiller IR 3 Compressed We air line S Vacuum line m Chiller tubing The chiller tubing provides the coupling fluid circulation path between the Echo liquid handler and the chiller The chiller maintains the fluid temperature at 22 C Tubing is sufficiently long to allow the chiller to be placed on the floor below the Echo liquid handler for reduced vib
125. nd one on the rear panel of the Echo liquid handler m To use the EMO switch in an emergency push it in a m To reset the EMO switch turn the knob clockwise until it pops out 3 2 8 Anti static bars The upper and lower anti static bars also called ionizer bars remove any electrostatic charge from the source and destination microplates Electrostatic charge can affect the placement of the transferred drop during liquid transfer 3 10 Echo Liquid Handler User Manual Figure 3 6 Anti static bars Upper anti static bar Lower anti static bars Barcode scanner The barcode scanner is optional You can purchase one or more barcode scanners to add to your Echo liquid handler that will read and report barcoded microplates Left Barcode Scanner Front Barcode Scanner not shown microplates You can use either the front scanner or the side scanner but not both at the same time System Description 3 11 O Note Several terms are used to specify barcode locations on a microplate Standing in front of an Echo liquid handler the two readable barcode locations are identified as follows Figure 3 1 Barcode label locations Left short or west position Front long or south position E Note Always place side barcode labels on the left side of both source and destination microplates The Echo liquid handler uses the left barcode scanner to read the barcode on the source plate but uses the right barcode scanner
126. nd side surfaces The obstructed areas are different depending on the microplate s flange height Refer to the ANSI SBS 3 2004 Bottom Outside Flange Dimensions specification for additional information B 1 B 1 Short flange height microplates Left side The barcode label may extend the full width of the microplate flat surface Its bottom edge may be no lower than either gt 1 0 mm above the top of the skirt or gt 3 0 mm above the bottom of the skirt Refer to Figure B 1 Front side Right side Back side Figure B 1 Short flange height barcode locations and obstructed regions Tee 21 0 mm above top of skirt gt 3 0 mm above bottom of skirt B 2 Medium flange height microplates Left side sides The 1 52 cm 0 6 in obstructed area is shown in Figure B 2 There is no obstruction on the front side of either source or destination microplates The right side of a microplate cannot be used for a barcode label The back side of a microplate cannot be used for a barcode label Figure B 2 Medium flange height barcode locations and obstructed regions 11 52 cml 6 62 cm 0 6in T 26n T e lObstructed x Barcode Scan gt area B 2 Echo Liquid Handler User Manual APPENDIX CHILLER INFORMATION The Labcyte chiller is a thermoelectric temperature control system that is included with your Echo system This chiller maintains constant temperature of the coupling fluid that is circulated
127. nged or the chiller is turned off C 4 Troubleshooting The Labcyte chiller has two system alarms that are shown on the chiller display RTD OPEN or RTD SHORT FAN FAIL RTD OPEN or RTD SHORT The connector has come loose or the temperature sensor has failed Turn off the chiller reconnect the cable and turn it back on If the alarm persists turn off and unplug the chiller and then contact Labcyte Service and Support See Contact information on page 7 1 FAN FAIL The fan is too slow to cool the chiller components or has stopped The fan is either blocked or has failed Check for obstructions to the side air inlet or outlet gratings of fan Ensure there is at least a 3 inch clearance around the fan If there are no obstructions to the air flow the fan may be failing Contact Labcyte Service and Support See Contact information on page 7 1 C 4 Echo Liquid Handler User Manual Labcyte Inc 1190 Borregas Avenue Sunnyvale CA 94089 1302 Telephone 1 408 747 2000 Toll free USA 1 877 742 6548 Fax 1 408 747 2010 email support labcyte com www labcyte com LABCYTE Echo Liquid Handler User Manual Labcyte Part Number 001 5331
128. nsing led to the application of focused acoustic drop on demand technology to life science liquid handling in the 2000s A 1 A 2 ADE and the Echo liquid handler The requirements of low nanoliter liquid handling in drug discovery and the biosciences are very exacting For the Echo liquid handler to transfer dissolved compound droplets while maintaining strict accuracy precision and throughput specifications the ultrasound acoustic pulse parameters must be adaptively calculated and the pulse must be precisely focused near the meniscus of the source microplate s fluid The primary variables that an Echo liquid handler must take into consideration when ejecting droplets include Volume of liquid to be transferred The transfer volume is specified in the transfer protocol definition Fluid composition Different fluid types exhibit different behaviors based on their surface tension interaction with microplate plastics and acoustic impedance tendency to attenuate sound waves passing through the fluid These differences determine the sound wave power timing and focus parameters required for successful ADE All Echo liquid handlers are pre calibrated to optimize these parameters for each fluid type Source microplate material and characteristics A transfer protocol s source microplate is specified in the transfer protocol definition from a list of Labcyte validated and predefined source plates Well fluid fill height The fluid f
129. o ensure that the droplets transferred from the source plate wells are directed upward into the center of the destination wells and not diverted to the side of the wells The grid should be inspected for particles and cleaned monthly Tool needed m Compressed air or vacuum with brush To clean the destination plate gripper grid 1 Display the Diagnostics window in the Echo software 2 Click Door Open to open the process door 3 Click Destination Plate Out The software extends the destination plate gripper and automatically turns off the electrical current Figure 6 1 Destination plate gripper extended 4 Gently remove any dust or particles on the grid with compressed air or vacuum with brush 5 Click Flip Destination Plate Down and repeat the previous step to clean the grid 6 Click Destination Plate In and select None from the Plate Name drop down menu in the Retract Dest Plate Gripper dialog box The software retracts the destination plate gripper 6 16 Echo Liquid Handler User Manual 6 2 7 Replace the AC power fuse The Echo main AC power entry module includes two user replaceable fuses Use only the correct specified fuse type as a replacement The Echo AC power configuration and replacement main AC power fuse are specified on the unit s product label on the rear panel Table 6 3 AC power configuration and fuses Echo AC power configuration Replacement AC power fuse 115 VAC 60 Hz 10 A 115 VAC 10 A FAS
130. oceed Figure 2 6 Reminder to close other applications Echo 5XX InstallShield Wizard xi Please close all running applications especially anti virus software Click OK to continue A dialog window will report the installation s activity and progress The installer must restart the client PC to continue installation 8 Click the Yes button Figure 2 7 Restarting the system EI Installing Echo 5XX The program features you selected are being installed Please wait while the InstallShield Wizard installs Echo 5XX This may take several minutes Status 1 Echo 5XX Installer Information xi configuration of Echo 5XX can continue Click Yes to A The installer must restart your system before restart now or No if you plan to restart later TnetallShield 9 Click the NEXT button after the workstation has restarted to continue the installation A dialog window will report the second phase of the installation s activity and progress This phase may take several minutes 10 Click the FINISH button when the installation is done 11 Remember to restart any applications you closed earlier such as the anti virus program 2 10 Echo Liquid Handler User Manual 2 3 2 Uninstall the Echo software If you need to uninstall the Echo software use the following procedure 1 Click the START button on the Windows Taskbar 2 Point to SETTINGS and select CONTROL PANEL 3 Double click ADD OR REMOVE PROG
131. ocol List displays the liquid transfer protocols that are stored in the Echo database TIP If you move the mouse pointer over a protocol name you will see a tool tip box that displays additional information about the protocol From the Protocols window you can click the appropriate button to perform the following tasks Add Click the Add button to create a new liquid transfer protocol Go to Creating a new a protocol for detailed instructions Edit Select the protocol name in the Protocol List and click the Edit button to change an existing liquid transfer protocol Go to Editing protocols for detailed instructions Remove Select the protocol name in the Protocol List and click the Remove button to remove an existing protocol from the Echo database A prompt box will ask you to confirm the protocol removal Run Select the protocol name in the Protocol List and click the Run button to perform a liquid transfer Go to Running a protocol for detailed instructions Click the icons at the top of the help page to navigate through the help pages BACK FORWARD HOME and to print the page OPTIONS gt PRINT help topics window Click the tabs in the left column CONTENT INDEX SEARCH to find specific Click the underlined text in the help page to jump to related subtopics Click the icon in the top left corner Fa and select CLOSE to close the HELP Echo Liquid Handler Software 5 1 5 4 Status window At the bottom of the
132. ode scanners The Echo liquid handler is considered as Class 1 laser light hazard under normal operating conditions with instrument covers on Emergency off label The Emergency Off label identifies the EMO Emergency Motion Off switch which is used in an emergency to shut down all mechanical Echo liquid handler activity There are two EMO switches one on the front panel and one on the rear panel of the Echo liquid handler Water system maintenance label LAB 5 YTE gt gt Regular water system maintenance required Flush system every two weeks Use distilled water only Do NOT use De lonized Water Use recommended water treatment See user manual for details The water system maintenance label is provided by Labcyte to remind the user to follow recommended maintenance procedures It also emphasizes the use of distilled water Echo Liquid Handler User Manual Pinch point label The pinch point label warns of possible injury if you do not keep your hands or Pinch Point fingers clear of source plate or destination Keep hands and microplate assemblies when they are fingers clear in motion High voltage label The high voltage label warns of possible high voltage injury from electrical cables High Vol connections and circuits inside the Echo gn vonage liquid handler Can cause serious injury Preface xvii Using this manual xviii Who should read this manual The Echo li
133. odically the instrument requests maintenance such as refilling the coupler fluid bottle emptying the waste bottle adjusting air pressure or fluid temperature Scheduled maintenance consists of purging the fluid system replacing the water filter washing the bottles and cleaning the anti static bars and destination plate gripper grid Six month preventive maintenance is provided by Labcyte field service representatives if the Echo liquid handler is still under warranty or under a service contract All user maintenance procedures are provided in this chapter This chapter contains the following sections m Maintenance schedule e Daily maintenance e Echo maintenance alerts e Scheduled maintenance m Maintenance procedures Refill the coupling fluid bottle Clean and refill the coupling fluid bottle Empty and clean the waste water bottle Replace the water filter Clean the anti static bars Clean the destination plate gripper grid Replace the AC power fuse m Extended Non Use and Storage e Maintenance During Extended Non Use Maintenance and Service 6 1 6 1 Maintenance schedule Echo user maintenance consists of the following m Daily maintenance m Echo maintenance alerts m Scheduled preventive maintenance e Every two weeks e Every month e Annual 6 1 1 Daily maintenance Perform the following checks once a day Table 6 1 Daily Maintenance Checklist Maintenance Task Procedure
134. of time has expired the Focus CALIBRATION indicator turns yellow The user should perform a focus calibration as soon as possible See Focus calibration check on page 5 57 Green Focus calibration has been performed recently Yellow A Focus calibration should be performed as soon as possible Coupling fluid temperature and indicator The coupling fluid temperature inside the Echo Coupling Fluid Temp 21 37 liquid handler is reported in degrees Celsius The RF Temp 4 43 45 C correct operating range for the coupling fluid temperature is 22 1 C 0 9 C The coupling fluid controls the temperature of the acoustic transducer A correct and stable acoustic transducer temperature is required to maintain accurate and precise liquid transfers The coupling fluid temperature indicator reports three states Green Coupling fluid temperature is OK Yellow Coupling fluid temperature warning The coupling fluid temperature is outside of the normal range 21 2 C 23 0 C Red Coupling fluid temperature fault The coupling fluid temperature is outside of the warning range 20 2 C 24 0 C RF Temp 1 temperature The temperature inside the Echo controller is reported in degrees Celsius The temperature sensor monitors one of the most critical internal electronic devices used to control the RF system Normal RF Temp 1 temperature is between 50 55 C As the reported temperature increases to 55 C or above it is possible that sourc
135. oftware 5 59 4 Click the SWEEP button to execute the focal sweep procedure The Echo acoustic transducer moves under the first well specified in WELLS and is moved vertically to the point where BB ToF equals the START TOF setting The focal sweep continues moving the acoustic transducer in Z INCREMENTS until a BB ToF equals the Stop TOF setting If multiple wells were selected the process repeats for each well Figure 5 1 Focal Sweep dialog box File Position vs Amplitude 3 73695 3 07867 2 42039 v Amplitude 1 76211 Start Stop ToF 1 10383 555 Plate Name 384PP_DMSO x 29 45 31 4266 33 4032 35 3798 37 3564 39 333 ToF us Z increment 50 um Display J Legend Wells Show M Placement Upper Left 5 After the impedance data is collected and analyzed it will be displayed in graphical form as shown in Figure 5 1 Which well data is viewed and which function s is plotted can be selected n the DispLaY section The plots are amplitude versus ToF BB Bottom of the Bottom The acoustic echo ToF returned from the interface of the bottom surface of the source plate bottom and the coupling fluid m TB1 Top of the Bottom the acoustic echo returned from the interface of the top surface of the source plate bottom and the well fluid TB1 BB Top of the Bottom Bottom of the Bottom The ratio between the TB1 echo amplitude and the BB echo amplitude m SR Surface Reflection
136. olds the source microplate containing the liquid that will be transferred For more information see Source plate gripper stage on page 3 5 3 2 Echo Liquid Handler User Manual Source plate insert The plate insert fits between the source plate and plate gripper to ensure that the source plate is properly positioned For more information and table of insert types see Source plate insert on page 3 6 Figure 3 2 Plate inserts Status indicator lights There are three status indicator lights on the front of the Echo liquid handler m Power green The Echo liquid handler is on and ready m Warning yellow A component is out of position or a parameter is out of range and requires attention m Fault red A problem has occurred that stops the current task and requires immediate attention These status indicator lights are also provided on the back panel For more information see Status indicator lights on page 3 8 LCD screen The LCD screen on the front of the Echo liquid handler reports what the Echo liquid handler is doing or what operator action is required This display is also provided on the back panel For more information see LCD screen on page 3 10 Emergency off The EMO Emergency Motion Off switch shuts down all mechanical Echo liquid handler activity This switch is also available on the rear panel Upper and lower anti static bars The upper and lower anti static bars remove an
137. om the source microplate to the destination microplate and in what well order For example a simple transfer protocol would transfer compound from source wells to the matching destination wells called picture perfect More complex protocols would transfer compound from different source plates to a single destination microplate The Echo liquid handler software provides a transfer map to specify the wells that will be used in the liquid transfer For a detailed discussion of different types of liquid transfer protocols see What types of liquid transfer are supported on page 1 11 Sample protocol parameters The example in the following procedure uses these parameters m Source microplate 384PP_DMSO 384 well polypropylene standard volume microplate for DMSO Destination microplate An empty 384 well microplate Transfer map The twenty wells defined as A1 through E4 Copies 5 Volume 10 nL transfer into each well Source well contents Sample fluid consisting of 10 mM compound of interest dissolved in DMSO 70 to 100 DMSO water 4 3 1 Add new protocol 1 From the PROTOCOLS window click the ADD button The New Protocol wizard will begin 2 Enter the protocol name for example 384 10 nL and select the source and destination microplate types from the drop down lists Figure 4 6 Entering protocol name Add New Protocol Wizard J Protocol Name 384 384100 Source Plate Type 384PP ER BP Destination Plate
138. ombinations To select an entire row or colur click the appropriate header To select all wells click corner header or press click the appropriate header To select all wells click corner header or press ctri A C Dest 1 AIS ITA TM On a gt iaai a Click the radio button for Dest 1 first destination plate Echo Liquid Handler Sofiware 5 21 b Select the wells you want to include in the transfer For an explanation on how to select wells in a transfer map see Create a transfer map on page 5 22 c Click the radio button for Dest 2 and select the wells you want to include in the transfer Repeat this process for the remaining destination plates O Note If you are selecting the same group of source wells for all four destination wells then click the radio button for All and select the wells you want to include in the transfer The software will automatically create the transfer maps for Dest 1 to Dest 4 6 Click the FINISH button after all four transfer maps have been created Create a transfer map A transfer map must be created for each source microplate The transfer map defines which source microplate wells contain compound that will be transferred to the destination plate When you click the View DEFINE button the DEFINE WELLS dialog box opens Figure 5 1 The Define Wells dialog box Define Wells Use your mouse to select the transfer wells To select more than one well use Control and Shift key comb
139. on their emitter pins and ground reference surfaces This accumulation could result in droplets being inaccurately ejected into the destination plate wells for example into the corners or onto the sides of a well Periodic cleaning will maintain proper anti static bar operation Once a month dry brush the anti static bars using the following procedure 1 Turn off the Echo carriage motors by pushing the EMO switch 2 Open the process door by clicking Door Open in the Diagnostic window 3 Turn off the anti static bars by clicking Anti Static Bars On in the Diagnostic window 4 Locate the two anti static bars See following figure Figure 6 1 Anti static bar location Upper anti static bar Lower anti static bar 5 Gently clean the emitter points and ground reference surfaces on both anti static bars with a dry soft bristle nylon toothbrush Figure 6 2 Anti static bar areas requiring cleaning Nine emitter points EE Two ground reference surfaces DO Note Leave the upper anti static bar in either its raised or lowered position either position is OK 6 Turn on the Echo carriage motors by turning the EMO switch counterclockwise and releasing it Maintenance and Service 6 15 Clean the destination plate gripper grid The grid on the destination plate gripper is charged with an electrical current during fluid transfer The electrical field overrides any static charge that may reside in the destination plate t
140. onding wells in the destination plate Figure 1 11 384 to 96 plate transfer 96 well 96 well Destination Microplate Destination Microplate 1 2 384 Well Destination Microplate P24 H12 113 gt A1 96 well Destination Microplate Destination Microplate 3 4 Introduction 1 13 1 3 3 384 plate to 1536 plate liquid transfer Figure 1 12 illustrates a 384 well to 1536 well interleaved fluid transfer An interleaved transfer places or stacks all source A1 well transfers into a single cluster on the destination plate Figures 1 13 and 1 12 clearly show these two schemes Each source microplate requires its own transfer map defining which wells are to be transferred to the destination microplate These transfer maps can be unique for each of the source plates If the transfer maps consist of single or multi region transfers the transfer protocol can be easily defined using the Echo software Complex transfer maps must be created programmatically using the Echo Activex programming interface refer to the Labcyte Echo Integration Guide Figure 1 12 384 to 1536 interleaved full plate transfer 384 Well 384 Well Source Microplate Source Microplate 1 2 1536 Well Destination Microplate 384 Well 384 Well Source Microplate Source Microplate 3 4 i 1 14 Echo Liquid Handler User Manual 1 3 4 1536 plate to 384 plate liquid transfer Figure 1 13 illustrates a 1536 well to 38
141. only as directed by Labcyte personnel 5 52 Echo Liquid Handler User Manual If register calibration is required use the following procedure 1 Click REGISTER in the Calibration window The Plate Registration window opens Figure 5 1 Plate registration window Plate Registration xi Plate Type Present Retract Register Labware Well Center Delta X 9 80 um Well Center Delta 7 22 um Select the source plate type from the drop down menu Click the PRESENT button to extend the source plate gripper stage Place the source plate on the plate gripper Click the RETRACT button to draw the plate into the instrument a uu A W N Click the REGISTER button The Echo liquid handler calibrates the well position and displays the new well center values 7 Click the APPLY LABWARE button at the bottom of the dialog box then Yes in the Confirmation Update dialog box to update the database Figure 5 2 Plate registration confirmed Fresenc KRecrace Register Labware Well Center Delta X 9 80 um Well Center Delta 7 22 um masso Confirm Update re you sure you want to update Center well for the 1536HB_DMS0 Plate Plate Skew Degrees _ _ Echo Liquid Handler Sofiware 5 53 5 8 2 Power calibration Proper acoustic droplet ejection relies on a calibrated RF energy source Environmental temperature changes can affect the characteristics of the Echo R
142. ontrols rEcho Liquid Handler Soft Reset Hard Reset reboot Shutdown 2 Leave the vacuum supply line connected and the vacuum source turned on 3 Switch off the AC power switch on the Echo liquid handler 4 Wait three minutes to allow the coupling fluid to completely drain out of the catch basin 5 Leave the chiller on and the air pressure and vacuum lines connected k Note Ifthe Echo liquid handler is going to be shut down for longer than a week start up and run the pump once a week to keep the acoustic transducer wet See Extended Non Use and Storage on page 6 18 4 4 Echo Liquid Handler User Manual EMO switch The EMO Emergency Motion Off switch can be used in an emergency to shut down mechanical Echo liquid handler activity There are two EMO switches one on the front panel and one on the rear panel of the Echo liquid handler To use the EMO switch in an emergency push it in When to use the EMO switch Any time the safety of the operator is in question Any time strange loud or potentially damaging noises are coming out of the Echo liquid handler When the instrument door will not open and the source or destination plate gripper stages collides with the door If water is leaking from the Echo liquid handler In any other situation that suggests immediate shutdown of the Echo liquid handler is appropriate Using the EMO switch The EMO switch performs the following actions A
143. ource or destination plate Fi gripper stage is extended outside of the Echo liquid handler when the process Xx ie o poce ae grippe e Daing Me doa door is commanded to close a Close ln Door Error error message is displayed The associated indicator lights report the current position of the door Echo Liquid Handler Software 5 31 Source plate in out and presence The source plate gripper stage can be commanded to extend outside or to retract back inside the Echo liquid handler Select the appropriate checkbox If the process door is closed when the source plate is commanded to extend outside of the Echo liquid handler the process door will be opened first When the source plate gripper stage is Retract Source Plate Gripper Ed commanded to retract into the Echo liquid handler the RETRACT SOURCE Plate Name 1536L0v_pmso x M PLATE GRIPPER dialog box will open SE i 1536HB_DMSO Select the plate type being loaded or 1536LDV_DMSO select NONE 1536LD _DMSO_HT a 384LD _AQ_B Two indicator lights are associated with Focus Calif 384LD _AQ_P the source plate One light reports the Temp 21 97 C sole current position in out of the source 43 45 C 384PP_DMSO plate gripper stage the other light 30D DMSO reports the presence of a microplate loaded onto the plate gripper The plate presence indicator has two states Clear Plate not present Green Plate present Destination plate in out and
144. ove RUN Introduction 1 7 1 2 4 System integration friendly While manual mode operator controlled via Labcyte software and stand alone mode hand loaded microplates support all functionalities the Echo liquid handler excels in its ease of integration into automated systems m The main body of the Echo liquid handler has a very small footprint 53 9 cm wide and 68 3 cm deep 21 2 in x 26 9 in m Facilities requirements power temperature ventilation compressed air vacuum communications are simple and straight forward m Source and destination microplates are loaded onto stages presented outside the instrument and placed in a fixed location for easy access by a robotic arm see Figure 1 6 m Most barcode standards are supported Barcodes can be located on the short or long side of a microplate m The Echo control software supports manual operation out of the box Echo device control system monitoring creation of destination plates and protocol definitions and protocol execution are available to the user m Customers choosing to develop their own Echo software applications are supported by a fully implemented ActiveX control library Figure 1 6 Robot loading a destination microplate laf n See What types of liquid transfer are supported on page 1 11 for more information 1 8 Echo Liquid Handler User Manual 1 2 5 Low maintenance requirements Echo instrument up time is high not only because
145. ower Calibrate Transducer Focal Sweep m Barcode Scanner Calibrate Echo Liquid Handler Software 5 51 5 8 1 Motion calibration Two Motion calibrations are available Home and Register Home calibration The HOME button moves each Echo stage destination plate gripper stage source plate gripper stage and acoustic transducer to the limits of its travel X Y Z or theta direction depending on the component to establish its absolute position m Destination plate gripper stage X left right Y front rear Z up down and theta plate rotation m Source plate gripper stage Y front rear m Acoustic transducer X left right Y front rear and Z up down During the homing procedure the destination plate gripper stage extends outside of the Echo liquid handler to rotate safely Figure 5 4 Homing the stages x Click on the Home button to home the stages cue After homing its stages the Echo controller knows the absolute position of each stage Register calibration The REGISTER button calibrates the well position for the selected source plate and updates the labware database This calibration is essential to position the acoustic transducer under the well center CS Note Normally the register calibration will not need to be performed by a user However it can be useful during certain repair diagnostic or recovery procedures Perform the register calibration
146. parameter of a user defined protocol can be edited however the destination plate type cannot be changed from 384 well to 1536 well or vice versa 5 24 Echo Liquid Handler User Manual 5 5 3 Remove an existing protocol definition If a protocol must be deleted select the protocol name and click the REMOVE button Confirmation of the removal is required The protocol definition is deleted from the Echo database and from the Echo software PROTOCOL LIST 5 5 4 Runa protocol To run a liquid transfer protocol 1 From the PROTOCOLS window select the protocol to be run and click the RUN button The first of a series of screens is displayed to provide instructions to run the selected protocol Figure 5 5 Protocol selected to be run Protocols Labware Diagnostics Calibration Advanced LABCYTE gt 384 10nL 5 copies 10 0nl 384 384 384 to 1536 1 copy 2 5nl 384 1536 I 384 to 384 1 copy 5 0nl 384 384 384PP Aqueous buff protein to 1536LDV 5 copies 2 5nl 384 1536 I 384PP DMSO to 384PS 1 copy 5000 0nl 384 384 Add Edit Remove Echo Liquid Handler Software 5 25 2 Click the NEXT button The process door will open and the source plate gripper stage will extend outside of the Echo liquid handler Run Protocol 384 10 nL 1 copy 10 0nl 384 384 Press Next to start the transfer The door will open and the source gripper will be presented 3 Load your source microplate with well A1
147. port the current rotational position of the destination plate gripper stage Plate dryer up down on off The Echo liquid handler can be commanded to raise or lower the plate dryer nozzle or to turn its vacuum supply on or off Select the appropriate checkboxes If necessary the Echo liquid handler will move the plate dryer beneath the source plate prior to raising it The associated indicator lights report the current position of the plate dryer and whether its vacuum is turned on or off Anti static bars on off The Echo liquid handler can be commanded to turn its anti static bars on or off at any time The associated indicator lights report whether the anti static bars are turned on or off Upper anti static bar up down The Echo liquid handler can be commanded to lower or raise its upper anti static bar The upper anti static bar can be raised or lowered regardless of the process door or source plate positions If the upper anti static bar is commanded to be lowered while the destination plate gripper stage is extended outside of the Echo liquid handler an Actuate lonizer Down Error will appear The associated indicator lights report on the position of the upper anti static bar Coupler nozzle up down The Echo liquid handler can be commanded to raise or lower its coupler fluid nozzle also called the bubbler at any time The associated indicator lights report the position of the coupler fluid nozzle Echo
148. presence The destination plate gripper stage can be commanded to extend outside or to retract back inside the Echo liquid handler Select the appropriate checkbox If the process door is closed when the destination plate is commanded to extend outside of the Echo liquid handler the door will be opened first When the destination plate gripper r stage is commanded to retract into the Echo liquid handler the RETRACT DEST PLATE GRIPPER dialog box will open Plate Name 384PP_Dest Select the plate type being loaded or Re pest _Des select NONE feaioy Dest ee 384PP Dest There are two indicator lights Focus Calif Greiner_1536HiBasePS_782096 associated with the destination plate o Sreiner_384P5_781096 One reports the current position in out 24 97 C None of the destination plate gripper stage The second reports the presence of a microplate loaded onto the plate gripper The plate presence indicator has two states Clear Plate not present Green Plate present 5 38 Echo Liquid Handler User Manual Flip destination plate The Echo liquid handler can be commanded to flip its destination plate gripper stage into the inverted or upright positions Select the appropriate checkbox If the destination plate gripper xl stage is still inside the Echo liquid handler when commanded to flip x eiO me ne a yoo ae upright a Flip Dest Plate Up Error will appear Lx I The associated indicator lights re
149. quid handler user manual supports several types of users involved in liquid handling and liquid transfer The Preface contains safety information that should be read by everyone Chapters 1 3 4 5 and Appendix A and B should be read by everyone Chapter 2 is of special interest to those involved in installing or relocating an Echo liquid handler Chapter 6 should be read by everyone who is responsible for user level maintenance and interfacing with Labcyte service and support personnel What is in this manual The Echo liquid handler user manual contains the following Chapter 1 Introduction discusses the Echo liquid handler its several operating modes and applications and how ADE acoustic droplet ejection is performed Chapter 2 Installation and Relocation covers pre installation requirements software installation and moving instructions Chapter 3 System Description describes the system components controls external connections status indicators and the LCD display Chapter 4 Manual Operation takes the reader through the performance of a sample fluid transfer protocol using the Echo liquid handler software Chapter 5 Echo Liquid Handler Software provides a thorough description of software functions to set up protocol and destination plate definitions run fluid transfer protocols perform diagnostic and calibration procedures and set up advanced fluid transfer options
150. r or from the bottom of the source microplates The inline water filter on the chiller tubing filters out dust and particles as the coupling fluid returns to the chiller To ensure that the coupler fluid is filtered effectively the water filter should be replaced monthly If the water filter appears discolored or cracked between monthly maintenance replace it immediately Figure 6 1 Water filter in good condition versus discolored Tools and materials needed m Inline water filter P N 001 4561 m Nylon hose clamps 11 32 in if needed P N 001 4045 m Compressed air or vacuum with brush To replace the water filter 1 Turn off the coupling fluid pump from the Diagnostics window 2 Remove the hose clamps from the ends of the water filter 3 Gently pull the water filter off the tubing 4 Insert the new water filter CS Note Ensure that the arrow on the water filter is pointing in the direction of the water flow away from the instrument 5 Secure the new water filter with the hose clamps 6 Turn on the coupling fluid pump 6 14 Echo Liquid Handler User Manual 6 2 6 Clean the anti static bars The Echo liquid handler uses two anti static bars to neutralize any static charge on the source and destination plates The anti static bars are just inside the process door one right below and the other right above the door opening Over time and use the anti static bars may accumulate dust or other contaminants
151. ration and good ventilation Compressed air line The compressed air line connects an external air supply to the Echo liquid handler Several of the Echo liquid handler mechanisms are pneumatically controlled These functions include opening the process door raising the acoustic transducer into position lowering the upper anti static bar and so on Air pressure must be between 80 to 150 psi m Vacuum line The vacuum line shown in Figure 3 1 connects the Echo liquid handler to a surge tank or directly to the vacuum supply The vacuum lines inside the Echo liquid handler pull coupler fluid back from the acoustic transducer and return the fluid to the coupler fluid bottle Another vacuum line removes coupler fluid from the bottom of the source plate just before it is unloaded Vacuum flow depends on the vacuum source house or pump See vacuum specifications on page 3 18 System Description 3 17 3 3 Specifications Each Echo liquid handler conforms to the following specifications 3 3 1 Physical Height 92 5 cm 36 4 in Width 53 9 cm 21 2 in Depth 68 3 cm 26 9 in Operating space envelope Maintain a minimum operating space of 59 cm 23 2 in width 99 cm 39 in depth and 118 cm 46 5 in height Position the Echo liquid handler in the operating space envelope to allow 2 5 cm 1 in side clearance add 17 8 cm 7 in if the coupling fluid bottle is placed on the side of the instrument 30 5 cm 12 in rear clearance
152. re Echo Liquid Handler Software 5 31 Barcode Location From the drop down list select the barcode location The choices are NONE LEFT SIDE and LONG SIDE front The right side is not supported CS Note Some users are more familiar with the microplate sides identified by the compass points LEFT SIDE west and LONG SIDE south Figure 5 1 Selecting barcode locations Pe ce eee FLBLOLEIBLE IESE EIEN le ad Left or west position Long front or south position 5 32 Echo Liquid Handler User Manual Plate dimensions Two illustrations identify the destination plate dimensions that must be entered These dimensions may already be specified in the SBS standards see SBS standards on page 5 35 Figure 5 2 Plate dimensions A B C and D A and B Well A1 X Y offsets Dimensions A and B locate the center of well A1 in relation to the plate s left and top outside edges A specifies the A1 well center to the left outside edge in millimeters B specifies the A1 well center to the top outside edge in millimeters Examples For a 384PP_DMSO plate type A is 12 13 mm and B is 8 99 mm The valid range for A is 0 00 mm to 128 00 mm The valid range for B is 0 00 to 86 00 mm Refer to ANSI SBS 4 2004 for more information C X center spacing Dimension C specifies the horizontal distance between well centers in millimeters That is the distance between wells A1 A2 A3 and so on Example For a 384PP_DMSO plate t
153. re can execute without triggering a no source plate loaded error message M Ignore Source Plate Sensor Check Ignore Destination Plate Sensor Check Default The source plate presence detect sensor is not ignored Ignore Destination Plate Sensor Check Selecting the IGNORE DESTINATION PLATE SENSOR CHECK checkbox bypasses the destination plate present sensor This may be useful when an individual destination plate does not fit correctly or droplets are being transferred to a microplate lid or other substrate that does not fit like a microplate A run protocol procedure can execute without triggering a no destination plate loaded error message Ignore Source Plate Sensor Check M Ignore Destination Plate Sensor Check Default The destination plate presence detect sensor is not ignored 5 9 2 Advanced solvent concentration DMSO only If the DMSO concentration in a source plate is known accuracy can be improved by specifying it prior to performing liquid transfers O Note The Solvent Concentration function assumes the solvent is DMSO therefore this function is not relevant for aqueous solutions The DMSO concentration information entered will be used with each source microplate until one of the following occurs m The USE PREDEFINED checkbox is cleared or m New DMSO concentration information is entered or m The Echo software is restarted Define Clicking the DEFINE m Solvent Concentration button open
154. re onto additional computers such as network PCs use the instructions provided in this section See Multi client considerations on page 1 21 for advantages and disadvantages of connecting more than one workstation DO Note Ifyou are connecting the Echo liquid handler to a LAN via a hub switch or router use a standard ethernet cable If you are connecting the Echo liquid handler directly to a workstation use a cross over Ethernet cable During installation the Echo software will provide the following installation types Figure 2 1 Installation types fe Echo 5XX Labcyte Inc x Welcome to the InstallShield Wizard for Echo 5XX The InstallShield R Wizard will install Echo 5XX on your computer To continue click Next Install Client Only f Stub Mode LABCYTE snstalserver on D C Py Install Client Server Normal Mode m Install Client Only Installs the Echo software onto the client PC that is connected to the Echo liquid handler normal mode The software allows the operator to communicate with control and monitor an Echo liquid handler through easy to use software screens Graphical User Interface This option requires a host name or IP address of the Echo server m Install Server Only Installs the Echo server software onto the Echo controller This option specifies the Echo model number and requires a license key The server software is installed by Labcyte field engin
155. roduct label on the next page for switch location See EMO switch on page 3 11 for a detailed explanation of what the EMO switch turns off when to use it how to use it and how to resume normal Echo liquid handler operation after using it xii Echo Liquid Handler User Manual Laser safety warnings The Echo liquid handler is a Class laser instrument that houses up to three optional laser based barcode scanners When operated as specified in this manual the Echo liquid handler does not expose the operator to laser light Nevertheless the Echo liquid handler contains a scanning barcode Class Il diode laser with maximum power up to 1 0 mW at 650 nm which is accessible in the interior of the unit The barcode scanners are not interlocked therefore if you operate the Echo liquid handler with the covers off you may be exposed to a Class II laser light hazard Do not operate the Echo liquid handler if its covers have been removed Preface xiii Product labels You will find the following labels on the Echo liquid handler 1 Product label ETL label Intellectual property label General warning label Laser safety label Emergency off label on EMO switches Water system maintenance label oe ER oe Pinch point labels source plate gripper stage destination plate gripper stage top and bottom 9 High voltage label destination plate gripper stage Figure 1 Echo liquid handler labe
156. roplate Droplets are ejected using an ultrasound transmitted from below the source microplate The ejected droplet is directly transferred upward from the supply reservoir to the receiver m No direct contact of the source fluid or the ejected droplet as it travels to the destination microplate means no cross contamination can occur Touchless fluid transfer provides the following benefits m Improved reliability m Lower maintenance requirements m Lower transfer costs m Reduced waste The following figure shows a single 5 nL droplet ejected from a 384 well microplate Figure 1 4 single droplet ejection Introduction 1 5 1 2 2 Reliability The Echo liquid handler has been designed for heavy duty cycles and reliable operation Compared to operating and maintaining existing liquid handling equipment the Echo touchless technology offers the customer a simpler trouble free device Proactive control and monitoring of the Echo systems reduce lost operational time or material if an error occurs m Multiple sensors monitor the coupling fluid system One sensor monitors the fluid level in the acoustic coupler catch basin Another sensor monitors for fluid at the low point of the Echo bottom pan m Compressed air and vacuum systems are monitored for correct supply and operation m Each of the moving mechanisms includes multiple sensors that define home positions and absolute limits reducing the possibility of mechan
157. rvey will be affected In turn inaccurate DMSO water ratio measurement can degrade the accuracy of fluid droplet ejection Calibrating the Echo acoustic transducer system compensates for certain changes in the transducer that can accumulate over time and use Focus calibration check A focus calibration check should be performed periodically The Echo liquid handler keeps track of how long it has been since the last focus calibration check was performed When a new focus calibration check should be performed the Focus CALIBRATION indicator on the Diagnostics tab will turn from green to yellow Figure 5 3 System Status section r System Status Power Air Pressure Warning Vacuum Pressure Fault Coupling Fluid EMO wasteBotte Fluid Leak Focus Calibration Coupling Fluid Temp 21 97 C RF Temp 1 43 45 C A Caution When the Focus CALIBRATION indicator is yellow Labcyte strongly recommends that the user execute the Focus calibration wizard as soon as possible Echo Liquid Handler Software 5 57 1 In the TRANSDUCER FOCUS CALIBRATION window click the Focus button Figure 5 1 Transducer focus calibration page 1 Transducer focus calibration step 1 of 2 rRange Current Focus ToF us 35 00 Start ToF us Stop ToF us 35 67 Z Increment um 50 7 Override Calculated Range Not Recommended Press Next to present source gripper and perform focal sweep 2 Accept al
158. s cass SA ns ns AR SS Ay D 1 9 1 2 7 Uncompromised accuracy precision and speed 1 10 1 2 8 Value added quality control 1 10 1 3 What types of liquid transfer are supported 1 11 1 3 1 384 plate to 384 plate liquid transfer 1 12 1 3 2 384 plate to 96 plate liquid transfer 1 13 1 3 3 384 plate to 1536 plate liquid transfer 1 14 1 3 4 1536 plate to 384 plate liquid transfer 1 15 1 3 5 1536 plate to 1536 plate liquid transfer 1 16 1 4 What system configurations are supported 1 17 Contents WI 1 4 1 Manual operation 1 17 1 4 2 Modular workstation operation 1 18 1 43 Small cell operation ss Lames Sax aw m ane dl aed 1 19 1 4 4 Fully automated operation 1 20 1 45 Multi client considerations 1 21 Chapter 2 Installation and Relocation 2 1 Pre installation requirements 2 2 2 1 1 Site requirements real wick Be eee Reda Re RES 2 2 2 1 2 Additional components 2 4 2 2 Installation overview soie tek aon a Raed a 2 5 2 3 Software installation 25242 Ada Beka ee kha ened chs ee mecha eels 2 6 2 3 1 Install the software ne Cuenca ees eee hed er ere wre re ar 2 7 2 3 2 Uninstall the Echo software 2 11 2 3 3 Upgrade the Echo software
159. s or command it to execute its functions m Uncoordinated clients could cause out of order execution that interferes with other users protocols For example client A could delete or edit a protocol or destination microplate definition created by client B An edited protocol definition would only be detected if the original definition was stored on client B and compared to the definition in the Echo database 1 22 Echo Liquid Handler User Manual CHAPTER INSTALLATION AND RELOCATION Labcyte installs the Echo liquid handler and Echo software for the customer However site preparation and additional software installation are the customer s responsibility This chapter covers pre installation requirements additional software installation and instrument relocation This chapter also provides a brief overview of the shipment contents and installation steps performed by Labcyte field engineers m Pre installation requirements Installation overview m Software installation Instrument Relocation Installation and Relocation 2 1 2 1 Pre installation requirements 2 1 1 Site requirements The following site requirements must be met before installation m Operating environment of 19 26 C 66 79 F and 10 80 relative humidity non condensing m Sturdy level water resistant bench top capable of supporting 128 kg 283 lbs with sufficient bench space to accommodate the Echo liquid handler
160. s presence with the source plate sensor Figure 5 3 Diagnostic survey plate map 7 Click the Go button The acoustic survey will be performed The acoustic transducer is moved in an efficient non stop path beneath each specified well Acoustic pulses are transmitted up into a well and the resulting return data is saved for analysis 8 After the survey the following options are Vi available in the View section below the es STATISTICS section Select the appropriate survey Volume type depending on the application read compestion Result Fluid Thickness e Volume Displays fluid volume e Fluid Composition Displays fluid composition in percentage DMSO water only e Fluid Thickness Displays well fluid thickness Post Processing e Raw The measured fluid percentages are not processed e Processed homogeneous The measured fluid percentages are processed using a moving average filter e Processed inhomogeneous The measured fluid percentages are processed to reduce measurement noise 3 The Echo 520 liquid handler does not report this result 4 Not available in Echo software version 2 2 and later 5 48 Echo Liquid Handler User Manual File Volume uL DIT A Results DMSO 70 100 Percent Volume See ae Fluid Composition Number of Rows 13 Fluid Thickness Post Processing Number of Columns 5 Riek Save Results to Disk V
161. s the Solvent Platemap a predefined Define dialog box Bee Echo Liquid Handler Software 5 65 Use predefined When the Use PREDEFINED checkbox is selected the Echo liquid handler will use the predefined DMSO concentration map Predefining a solvent concentration map CS Note Use this function carefully and only when you need to specify a fixed measurement and override the Echo DMSO measurement An incorrect DMSO concentration will produce questionable or unexpected results Place the cursor in any individual cell of the plate concentration map and enter the DMSO concentration for that well For example if the fluid contains 72 5 DMSO enter 72 5 Figure 5 1 Solvent platemap File Plate Name Sete 70 Yo Fill Solvent Concentration Percentage Enter a DMSO concentration in preparation for clicking the FILL button Plate Type Select the source microplate type Fill Clicking the FILL button loads the entered DMSO concentration percentage into each cell of the solvent concentration map File A predefined solvent platemap file in CSV format can be loaded by clicking FILE LOAD A predefined solvent platemap frie file in CSV format can be saved by clicking FILE SAVE Load r Save Ok Clicking the OK button closes the Solvent Platemap 5 dialog box and accepts the solvent concentrations entered Cancel Clicking the CANCEL button closes the Solvent Platemap dialog box with no action taken 5 6
162. semblies and reminds you to keep your hands and fingers clear of the mechanisms Pinch Point 3 2 2 Source plate gripper stage The source plate gripper stage is the lower plate stage that extends from the Echo liquid handler The plate gripper holds the microplate containing the liquid that will be transferred Note in Figure 3 5 that the source plate gripper stage extends out of the very bottom of the process door Figure 3 5 The extended source plate gripper stage Source plate gripper stage System Description 3 5 3 2 3 Source plate insert To keep the source plate secure the plate gripper includes a plate insert designed for specific plate types See the following table of plate inserts Table 3 1 Labcyte plate inserts Plate insert Plate catalog Insert size 384 well polypropylene P 05525 2 10 mm 384 well LV COC P 05710 1 50 mm 1536 well COC high base LP 03730 4 50 mm 384 well LDV COC LP 0200 4 50 mm 1536 well LDV COC LP 0400 4 50 mm CS Note Handle the plate inserts carefully They can be damaged from dropping or rough handling which can affect instrument performance When you are changing plate inserts look for the insert size which is located in the upper right corner When you place the microplate insert onto the plate gripper the tabs will be flush against the insert sensor The plate insert will rest on top of three pads and should move easily from side to
163. sfer volume directly instead of using the increment decrement buttons remember that the value must be an even multiple of the minimum transfer volume Echo Liquid Handler Software 5 17 3 Click the NEXT button to open the third ADD NEW PROTOCOL WIZARD dialog box Figure 5 3 Add new protocol wizard page 3 of 3 Add New Protocol Wizard Page 3 of 3 Source 1 Transfer Wels T viewDefine Source 2 Transfer Wels viewDefne Source 3 Transfer Wels view Define Source 4 Transfer Wels viewDefne You must define a fluid transfer map for each of the source microplates used 4 Click the View DEFINE button to create a transfer map for the first source plate The Define Wells dialog box shows the wells that are available for transfer Select the wells you want to include in the transfer Repeat this step for the other source plates and click the OK button Figure 5 4 Define wells in a transfer map Define Wells Use your mouse to select the transfer wells To select more than one well use Control and Shift key combinations To select an entire row or column dick the appropriate header To select all wells click corner header or press ctrl A For an explanation on how to select wells in a transfer map see Create a transfer map on page 5 22 5 Click the FINISH button after all four transfer maps have been created 5 18 Echo Liquid Handler User Manual Figure 5 5 384 1536 protocol finished Add New Protocol Wizard Page 3 of
164. side See Figure 3 1 Figure 3 1 Source plate gripper stage entire assembly Insert sensor Tab Pusher plate assembly Source plate insert Plate gripper gt 3 6 Echo Liquid Handler User Manual The plate insert will be moved into the correct position by the gripper arm when the source plate gripper stage moves through the process door When you load a source microplate onto the plate insert well A1 is located in the upper left corner see Figure 3 2 As a handy reminder the upper left corner of the plate insert is labeled A1 The insert contains tabs that inform the Echo liquid handler which plate is being used Figure 3 2 Plate insert and gripper arm securely hold the source microplate A1 well location _ Tabs 3 2 4 Destination plate gripper stage The destination plate gripper stage is the upper plate stage that extends from the Echo liquid handler The plate gripper holds the microplate that will receive the liquid being transferred When you load the plate onto the plate gripper well A1 goes into the upper left corner nearest the Echo liquid handler logo Figure 3 3 The extended destination plate gripper stage A1 well location When the destination plate gripper stage is commanded to retract into the Echo liquid handler the first action taken is to rotate the microplate upside down It is in System Description 3 7 this inverted position that the microplate will be able to re
165. so detect empty wells out of range fluid fill volumes or an incorrectly positioned acoustic transducer under a well wall A 5 A 4 Fluid transfer The Echo software performs an initial survey to determine the location of the meniscus and calculate the DMSO concentration for DMSO based solutions The software then uses dynamic real time measurements to check the meniscus position again and adjust the focus and power as necessary to compensate for any changes immediately before droplets are transferred from each well A4 1 Positioning the ADE elements Before a droplet can be ejected the source plate gripper stage and the acoustic transducer stage are moved on their X and Y axes so that the transducer is centered directly below the source microplate well from which fluid will be ejected The appropriate destination microplate well is positioned directly above the acoustic transducer and source microplate well The acoustic transducer assembly includes a fixed focal length lens In Figure A 3 below two adjacent wells with different fluid fill heights are shown Before a droplet can be ejected the acoustic transducer must be positioned vertically on its Z axis to correctly focus the acoustic energy near the fluid s meniscus Figure A 3 Fixed focal length dictates Z axis adjustment ee nd i ee A 4 2 Creating the acoustic pulse The acoustic pulse frequency amplitude and duration are calculated depending on the fluid
166. splash out Will the repeated motions of the destination plate or simply time passing allow any of the transferred liquid to fall or drain out of the destination plate well Testing at Labcyte under a variety of conditions shows that the following factors are responsible for eliminating splashing or other causes of fluid loss when the Echo liquid handler is used in liquid transfer applications m Relatively high surface tension of the transferred liquid m Low droplet velocity under 2 m s at deposition Pre loaded 1536 well destination microplates will not experience fluid loss when they are inverted Pre loaded 384 well destination microplates containing fluid with low surface tension may experience fluid loss when inverted and should be tested Pre loaded 96 well destination microplates may experience loss of fluid when inverted depending on fluid type and volume and are not recommended for wet pre loaded fluid transfers 1 4 Echo Liquid Handler User Manual 1 2 What are the Echo liquid handler features ADE and the Echo liquid handler are well suited to transferring low nanoliter volumes of aqueous and DMSO based solutions for a wide variety of applications Echo liquid handler features that will be important to any organization include 1 2 1 Touchless fluid transfers The Echo liquid handler uses a touchless technology m Nothing physically touches the fluid in the wells or the interior walls of the mic
167. tS x Sasa ee Mee yeni ee ete kre ay i ai ate erat Mer eel k C 2 C 2 Safety Warnings and Precautions C 2 C 3 Chiller Setup and Operation C 3 C3 SOUP ee Be be Lee te be eee Das ed ed C 3 3 2 Operation ame se ee Se nee ee Mia C 4 C 4 Troubleshooting a wre we Ne De C 4 Vi Echo Liquid Handler User Manual PREFACE The preface to the Labcyte Echo liquid handler user manual contains important information regarding the safe use of an Echo liquid handler and how to use this manual Read and understand the safety information thoroughly before you begin operating the Echo liquid handler m Intended audience m Safety warnings and precautions m Product labels m Using this manual The Labcyte Echo 520 550 and 555 systems are referred to as the Echo liquid handler throughout this manual The information and illustrations apply to all models unless specifically stated otherwise Intended audience The Echo liquid handler is designed to be used by individuals who are familiar with good laboratory practices GLP or similar laboratory safety program Echo liquid handler operators must read this manual before using the Echo liquid handler instrument Throughout this manual the word you or user refers to the Echo liquid handler operator 1 Good laboratory practice GLP for nonclinical laboratory studies FDA regulation 21 CFR Part 58 Preface vii Safety warnings and
168. tallation and Relocation 2 3 2 1 2 Additional components The following components are needed to work with the Echo liquid handler m Chiller for maintaining a constant fluid temperature Included with the Echo system See Chiller Information on page C 1 for more information m Distilled water for filling the chiller well coupling fluid bottle and coupling fluid circuit A Caution Do not use de ionized water as it may corrode the fluidics system m Client PC customer s computer running the Echo software to monitor and control the Echo liquid handler The PC should be an Intel Pentium III processor or equivalent with Windows XP Professional SP2 or later 256 MB of RAM minimum 200 MB of available disk storage video resolution of 1024 X 768 256 colors or better and a 10 100baseT Ethernet port amp Note If you are integrating the Echo liquid handler into an automated HTS High Throughput Screening system and need to install the Client Server software onto a workstation it must contain an Intel Pentium 4 processor or better m Network IP address A static IP address must be allocated for the Echo liquid handler The Echo liquid handler comes preconfigured with a static IP address 192 168 1 1 however if you cannot use this IP address in your network or if you are installing more than one Echo liquid handler contact Labcyte Service and Support to reconfigure the Echo liquid handler IP addresses See
169. tested to date This is attributed to the elimination of sample contact with the ejection mechanism In addition dynamic real time measurement of the fluid height at multiple times during a run allows the system to adjust automatically and instantaneously to changes in the fluid behavior over the course of the run interactions in the acoustic transfer process and the source liquid characterization performed during the source plate survey The use of a single high speed serial ADE mechanism reduces sample to sample variability with the Echo liquid handler as compared to tip based and other instruments that use multiple mechanisms in parallel 1 2 8 Value added quality control In addition to precise and accurate transfer of fluids the Echo liquid handler has an analytical mode that can provide quality control of fluid samples The Echo liquid handler applies a low energy sound pulse to the fluid sample in each microwell and the reflected signal is used to calculate the fill height and DMSO water concentration for DMSO based solutions The data can then be uploaded to a user database for post transfer processing and analysis For example Figure 1 7 shows part of the saved plate survey DMSO water concentration data CSV comma separated values file opened in Microsoft Excel Figure 1 7 An example of Echo DMSO water concentration data imported into a spreadsheet for analysis F1 Microsoft Excel 2004 02 12_09 32 38_platesurvey_DMS
170. the safety coordinator at your company for information A Safety cautions To protect the Echo liquid handler from damage follow these precautions Caution Always use the normal shutoff routine or the EMO Emergency Motion Off button to turn off the Echo instrument Turning off the instrument by other methods for example unplugging the power cord can potentially result in damage to the electronics or corrupt the software Caution Do not attempt to service or repair the internal Echo liquid handler mechanisms yourself The electrical laser and mechanical systems must be maintained by Labcyte field engineers If you have any questions regarding what may be serviced by an Echo liquid handler user please call Labcyte see Contact information on page 7 1 Caution Do not make modifications to the Echo liquid handler Caution Use only replacement parts that are approved by Labcyte Caution Use labware that is SBS compliant or approved by Labcyte Caution Do not obstruct the air vents Keep all air vents free of dirt or dust Caution Handle the Echo liquid handler coupling fluid and waste water carefully Avoid all spills 2 Society for Biomolecular Sciences See Supported labware on page 3 21 X Echo Liquid Handler User Manual Caution Keep corrosive agents or otherwise damaging material away from the Echo liquid handler and its attached devices Caution Do not expose the Echo liquid handler to ex
171. tions and system functions such as shutting down or restarting the instrument Function Keys Function keys perform tasks specific to the window in which they are displayed such as creating editing or removing a protocol Status window Two status windows are available Client and Server The Client status window shows the applications that are executed during a protocol run The Server status window shows the individual tasks that are performed within an application The status windows also display specific types of errors that the system encounters during a protocol run About and Help menu not shown Right clicking on a tab window background will display a menu that provides two choices About and Help About Select ABOUT to display the software version and build numbers Help Select HELP or press the F1 key to open the online Help file Each function window is described in more detail in the following sections About and Help windows Protocols window Labware window Diagnostics window Calibration window Advanced window Echo Liquid Handler Software 5 5 5 3 About and Help windows 5 3 1 About window You can display the ABouT window by right clicking on any tabbed window Protocols Labware Diagnostics Calibration or Advanced and selecting About in the menu Protocols Labware Diagnostics Calibration Advanced LABCYTE gt Protocol List 384 10nL 5 copies 10 0nl 384 384 384PP
172. types and use the naming convention platename_liquidtype_Dest such as 1536LDV_DMSO_Dest Destination plate types can be added edited or removed CS Note The plate list on your Echo system will vary based on the plate types you ordered with your system See Supported labware on page 3 21 Manual Operation 4 7 4 2 1 Edit the source microplate 1 Click the LABWARE tab in the Echo software to open the LABWARE window 2 Select 384PP_DMSO and click the Epit button 3 Select the barcode location from the drop down menu This the only parameter of a source plate that can be edited Figure 4 4 Source plate specification Plate Specification Manufacturer Labcyte Lot Number 04090130 Part Number P 05525 A1X Offset A A1Y Offset B R X Center Spacing C The source plate type cannot be removed from the labware list If you need to remove a source plate or add a different source plate contact Labcyte Service and Support see Contact information on page 7 1 4 8 Echo Liquid Handler User Manual 4 2 2 Define a new destination microplate If you are not using one of the pre defined destination microplates in the Echo software use the following procedure to define a new destination microplate 1 Click the LABWARE tab in the Echo software to open the LABWARE window 2 Click ADD to define a new destination plate type 3 Enter the new destination plate name required Optionall
173. uids that have been spilled on it m The Echo liquid handler or any of its subassemblies or components has been dropped or damaged m You suspect service or repair is required m The Echo liquid handler is being cleaned handled during a maintenance procedure or repaired except as directed in the applicable written procedure These conditions can cause an electrical shock hazard and result in severe injury Preface ix A Additional safety warnings Warning Do not try to gain access to the interior of the Echo liquid handler Do not remove the Echo liquid handler exterior covers Exposure to laser light high voltage or moving parts inside the Echo liquid handler can cause severe injury Warning Keep your hands fingers and clothing clear of the process door and microplate assemblies except when loading or unloading a plate Be aware of the possibility that either the source or the destination plate gripper stages may move Warning Use good laboratory practices and follow the manufacturer s precautions when working with chemicals Labcyte is not responsible or liable for any damages as a result of or as a consequence of the use of hazardous chemicals Warning The Echo liquid handler weighs approximately 128 kg 283 lbs Moving or lifting the Echo liquid handler incorrectly can cause severe injury Never attempt to move or lift the Echo liquid handler without using proper equipment and proper safety techniques Contact
174. ume 2 5 nL Maximum Transferred Volume DMSO Aqueous 384 Well PP polypropylene 10 uL 384 Well LDV low dead volume 5 pL 1536 Well LDV low dead volume 2 uL 1536 Well LDV HT low dead volume high throughput 2 uL 1536 Well HB high base 2 uL Transfer Resolution 2 5 nL 3 3 8 Supported labware 384 Well PP AQ BP aqueous w buffer and protein 10 pL 384 Well PP AQ SP aqueous w surfactant and protein 10 uL 384 Well LDV AQ B aqueous w buffer only 0 5 pL e 384 Well LDV AQ P aqueous w protein only 0 5 pL Source microplates The following Echo qualified source microplates are available Table 3 3 DMSO Source Plate Types Plate Calibration Vol range pL Working vol pL 384PP 384PP_DMSO 20 50 30 384LDV 384LDV_DMSO 2 5 12 9 5 1536HB 1536HB_DMSO 2 6 4 1536LDV 1536LDV_DMSO 1 0 5 5 45 1536LDV_DMSO_HT 1 HT High Throughput This calibration is available on Echo 555 models only It uses the same 1536LDV plate but with a faster transfer rate For transfer of higher volume or where precise droplet placement is not required see 1536 plate to 1536 plate liquid transfer on page 1 16 System Description 3 21 Table 3 4 Aqueous Source Plate Types Plate Calibration Vol range pL Working vol pL 384PP 384PP_AQ_ BP 20 50 buffer w protein 30 384PP_AQ_SP buffer w 20 50 surfactant amp protein 384LDV 384LDV_AQ B 3 12 9 buffer 384LDV_AQ_ BP 6 1
175. urce plate wide average from target volume Volumetric Precision lt 8 CV DMSO water concentration lt 8 accuracy error lt 5 precision CV Echo 550 and 555 models only Fluid temperature 21 C 5 C 70 F 9 F Compound should be in liquid form prior to transfer Throughput Transfer time for fluids in different plates and on different Echo systems Table 3 2 Fluid throughput comparison Plate type Transfer Echo 555 Echo 550 Echo 520 volume nL min min min 1 384PP_DMSO 2 5 1 4 2 5 2 5 2 384PP_AQ_BP 2 5 1 4 2 5 3 384PP_AQ_SP 2 5 1 7 2 7 6 1536LDV_DMSO 2 5 2 6 4 5 4 4 7 1536HB_DMSO 2 5 3 1 5 0 5 0 2 384PP_AQ_BP 2 7 38 3 384PP_AQ_SP 2 2 3 3 4 384LDV_DMSO 7 7 5 384LDV_AQ_B 6 1536LDV_DMSO 8 8 2 12 7 3 20 Echo Liquid Handler User Manual CS Notes 1 At low volumes transfer times for the Echo 520 and 550 systems are similar however as the volume increases the Echo 550 system shows a faster transfer time 2 Transfer time is dependent on the number of transfers per plate therefore for a full plate transfer times for a 384 well plate will always be faster than for 1536 well plate 3 At higher volumes the difference in transfer time between the Echo systems changes sig nificantly Supported fluids Can be calibrated for a wide range of aqueous and DMSO based solutions Echo 520 system supports DMSO based solutions only Minimum Transferred Vol
176. user interface software to define plate and protocol information The server is the Echo controller that runs the instrument motors and sensors Each subsystem is independent with its own system checks Introduction 1 21 This architecture allows the Echo liquid handler to be commanded by multiple clients to run concurrent operations yet ensure that each operation is safe with respect to each other For example one user might command the Echo liquid handler to do a plate survey then a liquid transfer protocol During the source plate survey another user might command the Echo liquid handler to extend the source plate The plate survey that is in progress will continue to completion then extend the source plate Thus while the Echo liquid handler guarantees safe operation it cannot prevent the commands of one user interfering with those of a second user Therefore it is the responsibility of the users to coordinate their actions to ensure the proper fulfillment of a transfer protocol Also if more than one user creates edits or deletes fluid transfer protocol definitions or destination microplate definitions then there must be communication between users to ensure that the definitions are consistent with the protocols Users should be aware of the following considerations with the Echo client server architecture m An Echo liquid handler is not limited to a single client One or more clients can either monitor its statu
177. utside of the Echo liquid handler 4 Load the destination microplate with well A1 in the inner left corner Click the Next button Figure 4 12 Loading destination plate Run Protocol 384 10nL 5 copies 10 0nl 384 384 Please load destination plate 1 of 5 Pressing Next will retract the destination gripper and the transfer will begin While the destination plate gripper stage inverts the microplate the source microplate is surveyed The Echo liquid handler determines the fluid composition for DMSO and well fluid height and then calculates the fluid volume in each well CS Note Fluid height for a given fluid volume can vary depending on the surface interaction of the fluid with the well walls Fluids that cling to microplate plastics will form steep menisci pulling fluid away from the center of the wells causing lower fluid height Fluids that interact less with the well walls will form shallow menisci leaving more fluid in the center and greater fluid height Therefore Echo may report different fluid heights for the same volume of different fluids 4 14 Echo Liquid Handler User Manual After the source plate survey is done and the destination microplate is positioned correctly above the source microplate the liquid transfer process begins Using the data contained in your destination microplate definition the protocol definition and the source microplate survey just performed droplets are ejected from each
178. vel 2 Set up the Echo liquid handler bottles computer and other ancillaries Remember to allow for minimum operating space around the Echo liquid handler See Site requirements on page 2 2 3 Set up and start the chiller See Chiller Setup and Operation in Appendix C for instructions 4 Set up the coupling fluid and waste bottles Ensure that the bottle caps are firmly attached to ensure a tight vacuum seal Place the filled coupling fluid bottle in the caddy The fluid level sensor red LED should be visible through hole in the top of the bottle caddy to confirm that the bottle is positioned correctly and is filled Figure 2 1 Fluid level sensor in bottle caddy Fluid level sensor O Note On newer systems the fluid level sensor red LED is on the side wall of the instrument Figure 2 1 Fluid level sensor in new bottle caddy Fluid level senso r Installation and Relocation 2 13 Figure 2 2 Bottle connections Coupler fluid bottle cap Waste bottle cap blue bottle cap red bottle cap To coupler catch nozzle From dryer nozzle From vacuum source From catch basin long straight tube J tube From vacuum Vacuum filter source 5 Connect the air pressure and vacuum pressure supply lines to the fluidics panel The connections are labeled for easy reference A Caution Do not run the pump until the vacuum source is connected and active Running the pump without vacuum can lead to floo
179. ves to the down position Coupler fluid nozzle moves to the down position Power lamp turns on green Warning lamp turns off yellow to clear Fault lamp turns off red to clear Coupler fluid pump turns on Power to the RF amplifier turns on Echo liquid handler warm up The Echo liquid handler requires a warm up period after the power is turned on The RF amplifier and the acoustic transducer must reach a stable operating temperature If the instrument has been off for an extended period of time a 60 minute warm up period is recommended to allow all components to reach proper operating temperatures CS Notes Echo liquid handler warm up time depends on the starting temperature of its internal components Always allow sufficient time for the Echo liquid handler to reach thermal stability Fluid survey or transfer precision and accuracy will be affected by using the Echo liquid handler before it is warmed up Manual Operation 4 3 41 2 Turn off Echo power You may need to power off the Echo liquid handler for any of the following reasons m Extended periods of non use m Preventive maintenance m Service or repair m Power interruptions such as scheduled facility power interruptions m Moving it to a new location When you need to turn the power off use the following procedure 1 Shutdown the controller inside the Echo liquid handler From the ADVANCED window click the SHUTDOWN button Figure 4 2 System reset and shutdown c
180. ward This step is required to receive droplets ejected UP from the source microplate See Will the fluid splash or fall out on page 1 4 1 2 Echo Liquid Handler User Manual Figure 1 2 Inverting the destination plate ot Loading destination microplate Plate being inverted Destination microplate down During the transfer process the source microplate remains in a fixed position the destination microplate is positioned just above the source microplate and moves simultaneously with the acoustic transducer The acoustic transducer moves beneath the wells At each well the acoustic transducer emits acoustic energy that ejects a specified volume of liquid upwards to the receiving well on the destination microplate Figure 1 3 Diagram of transferring fluid Transferred fluid n Destination plate Ejected droplet gt Source plate Fluid being transferred Acoustic transducer After liquid transfer the Echo liquid handler moves the filled destination microplate out of the instrument and inverts the microplate again so that it can be removed 3 The operator removes the destination microplate containing the transferred compound Introduction 1 3 1 1 2 Will the fluid splash or fall out The Echo liquid handler performs liquid transfer with an inverted destination microplate Users might question whether all of the droplet remains in the destination plate well after transfer Will any of it
181. y electrostatic charge from the source and destination microplates Electrostatic charge can affect the travel of the droplet For more information see Anti static bars on page 3 10 Barcode scanner option An Echo liquid handler can be purchased with one or more barcode scanners that will read and report barcoded microplates For more information see Barcode scanner on page 3 11 System Description 3 3 3 1 2 Echo liquid handler back panel Figure 3 3 Echo liquid handler back panel Status lights LCD screen LMA AC power and fuse compartment EMO switch ui Data connection Coupling fluid bottle and caddy Tubing and hose connections AC power and fuse compartment The AC power and fuse compartment also called the power input module contains the main Echo liquid handler power on off switch AC power outlet and fuse compartment For more information see AC power and fuse compartment on page 3 13 Waste water bottle on older Echo systems only The 500 mL waste water bottle collects overflow from the catch basin It is easily identified by its red cap For more information see Coupling fluid bottle on page 3 15 Coupling fluid bottle and caddy The 1000 mL coupling fluid bottle provides the coupling fluid distilled water that is pumped through the acoustic transducer to maintain constant temperature and transfer acoustic energy to the microwell It is easil
182. y enter the manufacturing information 4 Enter the number of rows and columns required A 384 well microplate has 16 rows and 24 columns Figure 4 5 Destination plate specification Plate Name fo Manufacturer LotNumber Part Number Rows fi Columns 24 Barcode Location Noe x A1X Offset a 1 mm ALY Offset 6 1 mm X Center Spacing C 4 5 mm Y Center Spacing D 45 mm Plate Height E 14 35 mm Flange Height F 2 41 m mm Well Width G 0 01 mm Well Capacity H 0 01 u 5 Select the barcode location if you are using barcode labels or select None 6 Enter the destination microplate A1 X and Y offsets X and Y center spacing plate height and well bottom width values required These measurements should be available from the manufacturer If not you will need to measure them 7 Enter the flange height and well capacity values You can use the default values These values are not currently supported and the data is not used 8 Click the OK button to store your new destination microplate definition To edit or remove the destination plate type select the plate type from the labware list and click the EDIT or REMOVE button CS Note For more information about defining the destination microplate see Manage labware definitions on page 5 31 Manual Operation 4 9 4 3 Create liquid transfer protocol The liquid transfer protocol specifies how much fluid will be transferred fr
183. y identified by its blue cap The coupling fluid bottle caddy contains a sensor that monitors the fluid level and issues a warning when the fluid level is less than 250 mL For more information see Coupling fluid bottle on page 3 15 Status light indicators LCD screen and Emergency off Same as the components on the front panel Data connection RJ 45 Ethernet network port connects the Echo liquid handler to a PC or to a network For more information see Data connection on page 3 14 Tubing and hose connections the Echo liquid handler uses tubing and hoses to connect to the chiller house air and house vacuum or surge tank and vacuum pump For more information see Fluidics panel on page 3 17 3 4 Echo Liquid Handler User Manual 3 2 Component description 3 2 1 Process door The process door is normally closed to protect the operator from moving mechanisms in the Echo liquid handler and from any stray light emitted by the barcode scanner lasers When a fluid transfer protocol is executed the process door will be opened automatically allowing easy human or robotic loading or unloading of microplates Figure 3 4 The process door closed Warning Any time the process door is open whether or not one of the microplate assemblies has been extended outside of the Echo liquid handler there is a pinch hazard The pinch point Kasp hands end label shown here is visible on both microplate fingers clear as
184. y motion off switch EMO xii Laser safety warnings xiii Product labels a ed a a ee xiv Product label asser ooh Veoh NEE EN NE E NT Son XV AME T RE ely ay he ROR aa Aad he TU BAe ee MAR XV Intellectual property label 220 x0 daw is ous ati eink XV Ele trical hazard o a riens eg a nC at a er ati r XV General warning label 22 sue si he et xvi Laser safety label succes sus a co rase Se eos xvi Emergency off label so a doa witty de vest da eet de strates xvi Water system maintenance label xvi Pinch point label xvii High voltage label onou a xvii Usingithis manuals sss esis 44 ponai menant SU Sekar ak o xviii Who should read this manual xviii What is in this manual xviii Conventions used in this manual xix 1 1 Whatis ADE A DC De CUS OUI dee Pa Je 1 2 1 1 1 How does the Echo system work 1 2 1 1 2 Will the fluid splash or fall out 1 4 1 2 What are the Echo liquid handler features 1 5 1 2 1 Touchless fluid transfers 1 sales 1 5 1 2 2 Reliability ee Be aed Dod BR NES DR ae BR ay wR 1 6 123 Esse OF USO as Sing Moa aR eA ARO RL REA ORS ES RAS 1 7 1 2 4 System integration friendly 1 8 1 2 5 Low maintenance requirements 1 9 1 26 Gost SAVINGS 5 i
185. ype C is 4 5 mm The valid range for C is 0 05 to 9 00 mm Refer to ANSI SBS 4 2004 for more information D Y center spacing Dimension D specifies the vertical distance between well centers in millimeters That is the distance between wells A1 B1 C1 and so on Example For a 384PP_DMSO plate type D is 4 5 mm The valid range for D is 0 05 to 9 00 mm Refer to ANSI SBS 4 2004 for more information Echo Liquid Handler Software 5 33 Figure 5 3 Plate dimensions E F G and H amit E Plate height Dimension E specifies the overall height of the plate in millimeters Example For a 384PP_DMSO plate type E is 14 4 mm The valid range for E is 6 50 to 16 mm LS Note When using barcode labels on higher plate heights for example 16 mm align the barcode label with the top of the plate to ensure proper scanning Refer to ANSI SBS 2 2004 for more information F Flange height Dimension F specifies the height of the plate s flange in millimeters The Echo liquid handler supports the SBS standard short medium and tall flange heights Select the correct one from the drop down list Example For a 384PP_DMSO plate type F is 2 41 mm The valid choices for F are 2 41 6 10 or 7 62 mm Refer to ANSI SBS 3 2004 for more information L Note The Echo liquid handler does not currently use the flange height property and any flange height dimension entry is ignored G Well bottom width Dimension G specifies

Echo 500 Series User Manual Software version 2.3

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