IN Cell™ Analyzer 1000 - GE Healthcare Life Sciences
Contents
1. iv Acquire fluorescence images 1 wavelength E WM Acquire brightfield image J Acquire phase contrast image M Acquire DIC image Cancel Finish Help Figure 4 11 Acquisition Protocol Wizard Microscopy IN Cell Analyzer 1000 Operating Instructions 28968377 AA 29 4 Operation 4 3 Defining an acquisition protocol If the camera has been set up with multiple configurations such as with different Gain or Binning factors select the configuration you wish to use Aquisition Hrotgeul izand Cameta Specify which camera pou are using from the drop down list A summary of camera parameters is displayed below Camera name CoolSnap HO Description Gaini 2 no binning Gair 2 Width 1300 Height 1026 Binning 1x1 Support OZ Module Yes 02 Module Image Quality Normal Flat Field Correction Mot established Use flat field correction if available Cancel lt lt Back Nert Figure 4 12 Acquisition Protocol Wizard Camera 9 Filter Selection and Display page is used to select excitation and emission filters for fluorescence studies and to specify a number of camera settings and parameters Monson Prutegh Wizard Filter Selection and Display Select the filters to be used from the list below Specify the exposure time offset and visuals for each filter Excitation filter Emission filter Exposure HWAF Visuals OZ Module nsec offse2. Figure 4 8 Acquisition Protocol Wizard Plate Heater Well Labels page is used to define a labelling scheme for each row and column of wells Poa l o 2 0 3 o ME 0 6 aterm Figure 4 9 Acquisition Protocol Wizard Well Labels 28 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Defining an acquisition protocol 4 3 7 Well definition Use the schematic diagram to designate wells as active or blank and to define the origin well i e the starting point for image acquisition By default all wells are initially designated as active isso Erotocol Wizard Well Definition Right click on the well positions to designate them as active or blank Images will be acquired from active wells only Set active selected well only O OO ae OO Set blank OO Set all but selected well acti ae et all but selected well active Set all active Set all blank OOO n OOO EEC OOOO OOOO 9 Cancel lt lt Back Next gt T me lds ds hele aS Se ee aes Figure 4 10 Acquisition Protocol Wizard Well Definition 8 Microscopy type If the transmitted light module has been fitted the fluorescence brightfield phase contrast or DIC imaging modes can be selected along with the required number of fluorescence imaging modes Microscopy Specify microscopy type and the number of wavelengths if applicable
3. WARNING A fume hood or similar ventilation system shall be used when flammable or noxious substances are used WARNING Fire Hazard Before starting the system make sure that there is no leakage WARNING Always use protective glasses and other personal protective equipment PPE appropriate with the current application to ensure personal safety during operation WARNING The operator should always follow proper laboratory procedures in handling and disposing of volatile or hazardous solutions IN Cell Analyzer 1000 Operating Instructions 28968377 AA 13 2 Safety instructions 2 1 Safety precautions Installing and moving the instrument WARNING Power cord Only use power cords delivered or approved by GE Healthcare WARNING Do not block the ventilation inlets or outlets on the system WARNING Access to power switch and power cord Do not block the rear and side panel of the instrument The Power switch must always be easy to access The power cord must always be easy to disconnect WARNING Laser Safety The Auto focus sensor should not be opened by the user or service technician Tamper proof screws have been employed on the case of the Auto focus sensor to prevent access to harmful invisible laser radiation Do not modify the Auto focus case or attempt to remove the tamper proof screws The laser employed in the Auto focus sensor is a 780 nm 5 mW laser WARNING Hot surfaces The microscope lamp houses a
4. IN CELL Analyzer 870 x 700 x 685 145 1000 Gen 1 1 Optical Unit Box 2 Service Cabinet 553 x 600 x 670 98 5 Box 3 Auxiliary 101 x 730 x 84 67 5 Components 3 3 Unpacking Contact your GE Healthcare representative upon delivery of the instrument Check the equipment for any apparent damage before allowing installation to start Document any damage and contact your local GE Healthcare representative NOTICE The IN Cell Analyzer 1000 instrument must be unpacked and installed by trained GE Healthcare personnel 3 4 Assembly Contact your GE Healthcare representative upon confirming receipt of the instrument components 20 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Installation 3 Connections 3 5 3 5 Connections A GE Healthcare representative will connect the instrument during the assembly process Figure 3 1 Service cabinet External Connections The unit has 2 main electrical inlets at the rear and 2 main power switches on the front The 3 connecting cables between the cabinet and the instrument may be connected in only one way as the cable ends are keyed to be accepted in only one of each of the three receptacles Also connected at the rear of the cabinet are the keyboard mouse and monitor second is optional and printer An earthed ground wire is installed Figure 3 2 Connections on the back of the instrument IN Cell Analyzer 1000 Operating Instructions 28968377 AA 21 3 I
5. Camera duplication Camera binning Cancel sequieitiun moro Wiza Specify which objective you are using from the drop down list A summary of objective parameters is displayed below NIKON 4 PLANAPO Ay 0 2 Back News o Help Figure 4 6 Acquisition Protocol Wizard Objective 5 Specify which microplate is to be used during image acquisition The Well Plate page of the wizard will list at least one plate type Aequiguy rumedl Wizard Well Plate Specify which plate to use from the drop down list A summary of plate parameters is displayed below Plate name Plate name Valid Humber of wells Well shape Plate height Top left well center Spacing Well size Bottom thickness Bottom height Bottom interace Well volume Cancel _ irish Greiner uClear Greiner uLlear es 96 8 s 12 Round 14 00 mm 14 38 11 24 mm 9 00 9 00 mm 6 50 mm 250 0 3 4 Plastic 350 0 Help Figure 4 7 Acquisition Protocol Wizard Well Plate IN Cell Analyzer 1000 Operating Instructions 28968377 AA 2 7 4 Operation 4 3 Defining an acquisition protocol 6 If your system is equipped with an optional plate heater the Plate Heater dialog box allows you to use plate heating during acquisition If plate heating is required check the Use plate heating box and specify the target temperature between 25 42 C Acquisition Protocol Wizard
6. Accuracy OB pm min stepping interval E of the objective depth of field M Show live image Cancel Finish Figure 4 21 Acquisition Protocol Wizard Software Auto Focus Search 16 Output Formats page of the Acquisition Protocol Wizard is used to specify the format of the output files Aeguisition Erotocol Wizard Output Formats Specify the format of the output images Press Help for more information Save images in proprietary im format Save images in TIF format lt lt Back Hest Finish Help Figure 4 22 Acquisition Protocol Wizard Output Formats 36 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Defining an acquisition protocol 4 3 17 Stack Destination page is used to specify where to store the images as they are acquired equity Protocol Wizard Stack Destination Specify base path where the image files are stored and the naming method for creation of subfolder for each plate System default Predefined C Determined at run time Image Stack Folder Naming Mode For every plate the system will create a subfolder of the base folder in which the image stack file will be stored Specify the method of naming these folders Use current date and time Example 2000 10 12 11 12 C Use the unique plate identifier Use a scrap folder Previous stack in this folder will be overwritten Cancel Finish Help Figure 4 23 Acquisition
7. Protocol Wizard Stack Destination 18 Batch Analysis allows you to analyze multiple image stacks i e images from different plates automatically with little or no intervention The Batch Analysis page of the wizard lets you put the image stack into the Batch Analysis Queue as soon as It has been acquired Bequisituuu Mrutueul ibe Batch Analysis Specify whether you want to automaticaly add acquired experiment to batch analysis QUueLIE iv Automatically add to Batch Analysis Queue Batch Analysis Queue file d apps sal33_releasequeue tet Analysis protocol Loo M Cancel lt lt Back Ment Finish Help Figure 4 24 Acquisition Protocol Wizard Batch Analysis IN Cell Analyzer 1000 Operating Instructions 28968377 AA 57 4 Operation 4 4 Preparations before start 19 Tocomplete the Acquisition Protocol Wizard press Finish As you exit the wizard the IN Cell Analyzer 1000 will re display the Acquisition Protocol Manager dialog box Asummary of the protocol is displayed Press OK to exit and save the protocol or press Run to run the protocol without going through the Start Acquisition Session Wizard 44 Preparations before start 1 Set the IN Cell Analyzer 1000 operation mode to Assay Development Mode gt Assay development 2 Create a Plate or Slide Definition from the Applications menu if one does not exist 3 Select the Open Plate Manager option from the option screen that appears when you ente
8. about 1 2 meters of normal bench space ca 70 cm wide This location should be away from strong drafts room temperature fluctuations intense sunlight vibrations and strong electrical or magnetic fields as may be produced by an NMR microwave oven large motors or refrigeration units In addition the mains power source 100 to 240 VAC should be properly grounded and free from voltage fluctuations harmonic distortions power dips and spikes The AC power line should be dedicated to the IN Cell Analyzer 1000 system and should not share that power with additional equipment Table 3 1 Power supply requirements AC Mains Requirements Specification Requirement Voltage 100 to 240 VAC Frequency 50 60 Hz It is emphasized that room temperature fluctuations i e maximum 2 5 C due to the cycling on off of heating and cooling systems strong air currents sunlight directly on the instrument and through space electromagnetic waves may cause subtle performance problems Table 3 2 Environmental operating requirements Environmental Operating requirements Temperature 10 to 40 C Humidity lt 70 Relative Humidity Atmospheric Pressure 700 hPa to 1060 hPa IN Cell Analyzer 1000 Operating Instructions 28968377 AA 19 3 Installation 3 2 Transport 5 2 Transport The system will be delivered in three separate crates the size and weight of each is given below Table 3 3 Crate sizes Crates Size mm Weight kg Box 1
9. GE Healthcare IN Cell Analyzer 1000 Operating Instructions Original Instructions Table of Contents Table of Contents EPRI een 5 11 Important USER N OrmMat ON s rscencsisscncemndearnaiisaiennmaianommataccies 6 1 2 Regulatory information sssssssssssssssssssssrsrrisssssssssssssssrsssrsririrressssssnsssnsssrrrerrrrress T LS i a Uae gt gh onion a a 9 2 Safety MIS UUICTIONS saison 11 21 Safety procoutionS sanesna a N 11 B O eon E EEEE E T S 16 23 Recycling POCO SS i sahcccrisccrseesncssaasabsncaSeaiavsiatdetintsnnautanauutnntdee 18 3 Installation saniinacccanpecesnsisornerstnstnsvciesensseasavensineeasbentannelcoranveieskel 19 ad MTN O N a A T TAA 19 D TTN a RTT NRT En SMO RICME Ce MOAI Oreck CORt mT AT A maT oe ONT eT ee 20 Goo UOC a ters recente enren EE E A ETAS 20 34 SASS CMO sirsin destenncscutietausse tert eacsn aed cated datacenter suaaee 20 cs CA 6 0 g aE 21 3 6 Spare parts and ACCESSOLIES waeceeecsessssssssessseessssssssecsseccssecssecsssecssecsssecesecesseesses ec A MOS POLUON ena 23 41 Operation OVEIVICW ciartncntnminictiecannnmmensmanananenne 25 4 2 Starting the instrument oo eeecescsesssessssesssseesseccssecssessssecsssssssecsssccssecsssesssecsseeessess 24 4 3 Defining an acquisition protocol ssssssssssssssssssssiiirrisssrsssssssssssssssrsrrrrrrsss 25 4 4 Preparations before start ssssssssssssssssisrissssssssssssssssssesrrrrrrssssssssssnsssrrrrrrrrrnns 38 4 5 Acquirin
10. IN Cell Analyzer 1000 instrument contains a non operator replaceable fuse Power failure IN Cell Analyzer 1000 1 Close the IN Cell Analyzer 1000 software and shut down the computer 2 Switch off the lamp power supply and then the instrument Remove all mains cables from the power outlets 2 3 Recycling procedures The equipment shall be decontaminated before decommissioning and all local regulations shall be followed with regard to scrapping of the equipment Disposal general instructions When taking the IN Cell Analyzer 1000 system out of service the different materials must be separated and recycled according to national and local environmental regulations Recycling of hazardous substances The IN Cell Analyzer 1000 instrument contains hazardous substances Detailed information is available from your GE Healthcare representative Disposal of electrical components Waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately Please contact an authorized representative of GE Healthcare for information concerning the decommissioning of equipment i 18 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Installation 3 Site requirements 3 1 3 Installation NOTICE The IN Cell Analyzer 1000 instrument must be installed by trained GE Healthcare personnel 3 1 Site requirements The IN Cell Analyzer 1000 with Computer Controller requires
11. are except for alterations described in the user documentation or explicitly authorized by GE Healthcare Regulatory compliance of connected equipment Any equipment connected to the IN Cell Analyzer 1000 should meet the safety requirements of EN 61010 1 IEC61010 1 or relevant harmonized standards Within the European Union connected equipment must be CE marked 8 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Introduction 1 Instrument 1 3 1 53 Instrument Figure 1 1 The Services Cabinet contains the following Computer Lamp power supply unit 24 volt DC power supply unit Main power service rail Power cabling connections Liquid handling power supply Kinetics option OZ Amplifier OZ option Ludl encoder control box The optical unit contains the following Inverted microscope and base plate IN Cell Analyzer 1000 Operating Instructions 28968377 AA 9 it La 10 Introduction Instrument e Plate delivery mechanism 1 Figure 1 2 Plate delivery system 1 e Objective nosepiece e Auto focus and stepper motor e Liquid handling mechanism Kinetics option e Heaters Kinetics option e Optigrid paddle OZ Option Xenon Lamp Assembly This is a high intensity light source used for microscope observations only It should not be used for any other purpose Do not disassemble the lamp as this will affect the performance of the instrument To replace the bulb contact a GE Healthcare
12. are the property of their respective owners 9184 U ppsa la 2010 General Electric Company All rights reserved Sweden First published Jan 2010 All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them A copy of these www gel ifesciences com terms and conditions is available on request Contact your local GE Healthcare representative for the most current information GE Healthcare UK Ltd Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare Japan Corporation Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan imagination at work 28968377 AA 01 2010
13. ce please visit www gelifesciences com contact or for IN Cell Analyzer 1000 specific information visit www gelifesciences com Problems that prevent users from operating the instrument require immediate consultation with a GE Healthcare technician Customers should not attempt to repair the hardware or software unless instructed to do so by a GE Healthcare service representative IN Cell Analyzer 1000 Operating Instructions 28968377 AA 51 6 Troubleshooting 6 1 How to get help for IN Cell instruments 52 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Reference information 7 Instrument specifications 7 1 7 Reference information 7 1 Instrument specifications General specifications Part Characteristic Data Camera Pixel size 6 45 um x 6 45 um Field of view at 10 x 0 603 mm Pixel layout 1392 x 1040 Focus Method Confocal laser sensor Objective positioner resolution 0 2 um piezzo Objective resolution Illumination Epiflourescent lamp Transmitted light Magnification 4 x 0 20 Plan apo 10 x 0 45 Plan apo 20 x 0 45 ELWD Plan Fluor 40 x 0 60 ELWD Plan Fluor Plate positioning Resolution IAY etage Speed well to well on a 96 well u plate Filter selection Excitation filters Emission filters IN Cell Analyzer 1000 Operating Instructions 28968377 AA 53 7 Reference information 7 1 Instrument specifications Physical specifications Description Doh Ke Instrument 700 x 870 x 685 mm dimensions Serv
14. ck dialog box displays a color coded schematic diagram of the microplate and all the well locations are labelled Click Start to begin the acquisition process tag excluded from acq n no images acquired acq n in progress acquired 4 Image wel ME Fe o time 7 z J Autocontrast Mam af Linked exposures Path D Apps 4L33_Shipping IMGSTACK Hello World d Done Help View thumbnails Start Figure 4 33 Acquire stack window Optical Z Sectioning Module optional The optical z sectioning module allows you to convert the illumination system of a conventional wide field microscope to a structured light imaging system Before taking optical z sectioning images the optical z sectioning module must be calibrated The steps are as follows 1 Dock the plate 2 Insert the optical z sectioning calibration slide into the IN Cell Analyzer 1000 Select Application gt Dock Plate The dock plate slides out A window appears with the following message Put the plate on the plate holder Press OK when ready Place the calibration slide part of the optical z sectioning package in a slide holder and insert the slide holder onto the plate holder Click OK The dock plate slides in 3 Bring the optical grid pattern into focus 44 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Acquiring images 4 5 Select Settings gt Calibrate OZ Module The follo
15. ders equisitigy Protocol izana f Stack Destination Specify base path where the image files are stored and the naming method for creation of subfolder for each plate System default D Sppss4 33 Release IMGSTACKACopy of ha Predefined _ Determined at run time Image Stack Folder Naming Mode For every plate the system will create a subfolder of the base folder in which the image stack file will be stored Specify the method of naming these folders Use current date and time Example 2000 10 12 11 12 C Use the unique plate identifier Use a scrap folder Previous stack in this folder will be overwritten Cancel Finish Help Figure 4 30 Acquisition Protocol Wizard Stack Destination 42 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Acquiring images 4 5 4 Plate Layout Information dialog box allows you to preselect wells to be acquired Select a layout file which contains the configuration for the acquisition Figure 4 31 Acquisition Session Wizard Plate Layout Information 5 Instrument Preparation page displays Start Acquisition Session f r different set of filters Figure 4 32 Acquisition Session Wizard Instrument Preparation e Specified objective IN Cell Analyzer 1000 Operating Instructions 28968377 AA 43 4 Operation 4 5 Acquiring images e Flat field Correction e Plate Heating 6 The Acquire Sta
16. e well select which field to display from the Field drop down list If images were acquired at multiple wavelengths select which wavelength image to display from the drop down list 3 Manually digitized images may be saved to disk as stand alone image files in TIFF tif or proprietary MCID im format Individual stack images may also be stored as discrete files in the same manner 4 7 Procedures after a run 1 Close the IN Cell Analyzer 1000 software and shut down the computer 2 Switch off the lamp power supply and then the instrument Remove all mains cables from the power outlets 3 Microplates and any used materials in the instrument must be disposed of in the manner prescribed by local standard operating procedures There are no other waste materials resulting from the use of the IN Cell Analyzer 1000 CAUTION Hazardous waste must be specially handled and disposed of properly IN Cell Analyzer 1000 Operating Instructions 28968377 AA 47 4 Operation 4 7 Procedures after a run 48 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Maintenance 5 Daily checks and routine maintenance 5 1 5 Maintenance This section provides the user with information on the proper maintenance of the instrument to ensure proper function The maintenance methods described below do not require working anywhere near the exposed hazard areas 5 1 Daily checks and routine maintenance The following should be checked performed o
17. fore using the system Do not open any covers or replace parts unless specifically stated in the user manual The system complies with applicable European directives Labels concerning hazardous substances This symbol indicates that the waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately Please contact and authorized of the manufacturer for information concerning the decommissioning of equipment This symbol indicates that the product contains hazardous materials in excess of the limits established by the Chinese standard SJ T11363 2006 Requirements for Concentration Limits for certain Hazardous Substances in Electronics Emergency procedures In an emergency situation do as follows to stop the run Step Action 1 Disconnect the equipment from the power outlet f Check that the mains power source is providing power by connecting another electrical item to the power outlet If the mains power source does not work contact your local maintenance department before proceeding E Check that the mains power cable is connected correctly and that the plugs are firmly located in the sockets IN Cell Analyzer 1000 Operating Instructions 28968377 AA 17 2 Safety instructions 2 3 Recycling procedures Step Action 4 Test the unit again by switching it on 5 If the unit still does not work contact your local GE Healthcare service engineer because the
18. g wash the system with a neutral solution in the last phase or step WARNING Decontaminate the equipment before decommissioning to ensure the removal of all hazardous residues IN Cell Analyzer 1000 Operating Instructions 28968377 AA 15 2 Safety instructions 2 2 Labels 2 2 Labels Labels on the instrument Below are examples of the identification labels attached to the IN Cell instrument XX XXXX XX INCell Analyzer xxxx Code No XXXXXXXX Voltage Serial No XXXXXXX Frequency Mfg Year 2009 Power max Fuse A g D C UUM TI N12406 Made in Sweden GE Healthcare Bio Sciences AB 751 84 Uppsala Sweden Figure 2 1 Marking label CLASS 1 LASER PRODUCT Per IEC 60825 Figure 2 2 Laser warning label Laser Auto focus The IN Cell Analyzer 1000 uses a Class laser guided Auto focus device The laser is contained within the Auto focus device which is integrated with the microscope nosepiece WARNING The Auto focus sensor should not be opened by the user or service technician Tamper proof screws have been employed on the case of the Auto focus sensor to prevent access to harmful invisible laser radiation 16 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Safety instructions 2 Labels 2 2 Symbols used in safety labels The system complies with the requirements for electromagnetic compliance EMC in Australia and New Zealand Warning Read the user manual be
19. g IMAG SS ticsssasrensmiserancaiaccanennmiienuuaiiomsaniananuaanuatnmedan 40 46 Displaying animMage STACK icsseieassasivescrnevsiseantartnesrmasviommtanvndemcaesvenien 46 4 7 Procedures ofter A rUn s sisssssevsossiniarealsinivtasersnsnncinnsytnlaute annetiaascersartuvosidels 47 5 MaMia E ernn 49 5 1 Daily checks and routine MAINTENANCE sssssssssssessssessssrirrissssssssssssrssrenn 49 5 2 Component MAINTENANCE ssssss ssssssssssessssssssssssssrirrrssssssrsssssssssrssrsrirrrsreerssrssssssss 50 6 Troubleshooting sirina 51 6 1 Howto get help for IN Cell instruments sssssssssssssssssssissssssssssssssssrsrsrrrrress 51 7 Reference information scsscsssscsscssecssessecscessesseesseeees 53 7 1 Instrument specifications sasccisscssssssssasasvinsntusiavesssssesuesendevineslaseddvessunsisdssendavivasen 53 Le Ordening TNO ICO cease tssansee inden estes ussnconissnnescatiosesesieaueivortaanionaunionaiiaenaninnt 55 IN Cell Analyzer 1000 Operating Instructions 28968377 AA 5 Table of Contents IN Cell Analyzer 1000 Operating Instructions 28968377 AA Introduction 1 1 Introduction Purpose of the Operating Instructions The Operating Instructions provide you with the instructions needed to handle the IN Cell Analyzer 1000 in a safe way Prerequisites In order to operate the IN Cell Analyzer 1000 safely and according to the intended purpose the following prerequisites must be met e You should have a general understanding of the
20. ice cabinet 600 x 553 x 670 mm dimensions Environmental 280 x 140 x 185 mm control module dimensions Instrument weight 125 kg Service cabinet 85 kg weight Environmental 3 5 kg control module weight For operation the IN Cell Analyzer 1000 with a computer controller requires about 1 2 meters of normal bench space ca 70 cm wide 54 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Reference information 7 Ordering information 7 2 Electrical Specifications Characteristic IN Cell 1000 Voltage 100 to 240 Volts AC Frequency 50 to 60 Hz Power 70 Watts Output Secondary Data connection only Protective Earth Internal external marking Terminals Environmental conditions Condition Characteristic Limits Humidity 20 to 60 RH 7 2 Ordering information For ordering information visit www gelifesciences com IN Cell Analyzer 1000 Operating Instructions 28968377 AA 55 7 Reference information 7 2 Ordering information 56 IN Cell Analyzer 1000 Operating Instructions 28968377 AA For local office contact information visit ce imagination at work and GE monogram are trademarks of General f Electric Company www gelifesciences com contact IN Cell is a trademark of GE Healthcare companies Any use of this software is subject to GE Healthcare Standard Software End GE Healthcare Bio Sciences AB User License Agreement for Life Sciences Software Products Bjorkgata n 50 All third party trademarks
21. ion Protocol Wizard Liquid Handling 11 Z dimension feature Figure 4 16 allows you to collect a set of images at different focal planes Check the Enable acquisition of images in Z dimension box and enter the required Number of Slices images the Starting position and the Step size in the corresponding fields Up to 50 slices are available and the starting position can be between 100 and 100 microns from the HWAF position Slee paige epee ye Fer Ben ST Figure 4 16 Acquisition Protocol Wizard Z dimension 52 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Defining an acquisition protocol 4 3 12 Acquisition Options page is used to specify the manner in which the images are to be acquired The schematic Well Layout diagram provides a representation of the entire well and the arrangements of the field of view s selected Acquisition Protocol Wizard Acquisition Options Specity how mages are to be acquired Press Help for more information Flexible number of fields Acquire until l cells are found Fields to be acquired i ange liek Direction of Acquisition Horizontal Vertical Well Layout i Horizontal Snake C Vertical Snake Acgui ition Pattern f Acquire at well center C Acquire at a epecified location Acquire randomly in specified active region is Cente FE Er Edit coves eak Nee tien ee Figure 4 17 Acquisition Protocol Wizard Acquisition Opti
22. n a regular daily basis Data Back Up It is recommended that you back up the data files and programs in the operating computer in accordance with procedures set up by your local data administrator Environmental Checks Check that the room where IN Cell Analyzer 1000 is being used fulfils the environmental requirements listed in Section Environmental conditions on page 55 Computerized Online Diagnostics Diagnostic checks of the stage liquid dispenser filter wheels shutter and Auto focus mechanism are accessed from the Application gt Troubleshooting gt Self diagnostic Test menu Select the required components to test and click OK The results are saved to file in the Session logs folder of the IN Cell Analyzer 1000 software Cleaning the System Do not try to clean inside any of the component parts of the instrument Clean off any dust and dirt from the outside of the instrument using a lint free cloth slightly moistened but not soaked with 70 ethanol Wait until all parts are dry CAUTION A N If any parts of the system should become wet immediately close down all parts of the system and disconnect the power cables from the mains outlets Wipe the wet parts with a lens tissue or lint free cloth IN Cell Analyzer 1000 Operating Instructions 28968377 AA 49 5 Maintenance 5 2 Component maintenance 5 2 Component maintenance Stuck Charge High sensitivity CCDs such as the one used in the IN Cell Analyzer 1000 came
23. nd its surroundings become extremely hot during lamp illumination Do not touch any of these surfaces while the lamp is lit or immediately after the lamp has been extinguished CAUTION Pinch hazard Your fingers could be pinched if they are near the stage when it is in motion CAUTION Waste tubes and containers shall be secured and sealed to prevent accidental spillage 14 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Safety instructions 2 Safety precautions 2 1 Maintenance WARNING Replace fuses ONLY with same type fuses Several spare fuses are provided with the original shipment and the power receptacle is labeled with the correct type WARNING Repairs alterations or modifications must only be carried out by a GE Healthcare specialist or with explicit directions from a GE Healthcare technician Removal or modification of any cover or component could result in an unsafe or easily damaged instrument The GE Healthcare service department will be happy to answer any questions and provide parts and service when necessary WARNING Only spare parts that are approved or supplied by GE Healthcare may be used for maintaining or servicing the system WARNING Disconnect power Always disconnect power from the instrument before replacing any component on the instrument unless stated otherwise in the user documentation WARNING Hazardous chemicals during maintenance When using hazardous chemicals for cleanin
24. ng IMGS TACK Hello World A A xdce C Open an existing image stack file ooo Cancel lt lt Back Finish Help Figure 4 28 Acquisition Session Wizard Acquisition e Create a new image stack file Select to create a completely new image stack e Continue with current image stack file Select to continue with the stack that is already open filename is shown e Open an existing image stack file Select to open an existing stack Press Browse to locate the stack file IN Cell Analyzer 1000 Operating Instructions 28968377 AA 41 4 Operation 4 5 Acquiring images 2 Select the appropriate Acquisition Protocol from the Protocol name drop down menu Start Acquisition Session g Select Protocol Select a protocol from the drop down list A summary of the protocol s parameters is displayed below Protocol name Protocol name default Microscopy type Fluorescence File name None Plate Greiner uClear Camera CoolSnap HO Objective Nikon_10x_Plan4po Fields per well 4 2 2 Auto focus search Off Combining Off Direction Horizontal Snake Number of active wells 96 Number of acquired images 0 of 384 Time series Off Expected running time Off Cancel Finish Help Figure 4 29 Acquisition Session Wizard Select Protocol 3 Stack Destination page is used to specify where to store the images as they are acquired and to specify a method for naming the fol
25. nstallation 3 6 Spare parts and accessories 3 6 Spare parts and accessories For correct up to date information on spare parts and accessories visit www gelifesciences com 22 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Operation overview 4 1 4 Operation 4 1 Operation overview Step Action 1 Switch the instrument on 2 Ensure any manually adjustable hardware is in place 5 Select or create an Acquisition Protocol 4 Acquire an image stack 5 Select or create an analysis protocol 6 Load and analyze the image stack CAUTION Before using the IN Cell Analyzer 1000 ensure that the system is clean and dry Any detected problems should be dealt with before the system is started up If there is a spill or some of the modules have condensed water these should be dried clean and the system left for a few hours before using IN Cell Analyzer 1000 Operating Instructions 28968377 AA 23 4 Operation 4 2 Starting the instrument 4 2 Starting the instrument 1 Switch on the instrument using the On Off switches in the Service Cabinet L von Sy sae a LLII Ld IN Cell Analyzer 1000 46 seeeeeeeeeseseeseeeeenee Figure 4 1 Service cabinet closed 2 Ensure that the dichroic is correct type and position 3 Ensure that the required objective lens is correctly positioned by turning the turret until you feel it click into place 4 Switch on the lamp power supply by turning the key cl
26. ockwise Warm up time is 15 min F f f f j F j L i d d I Sounn Figure 4 2 Key used to switch on lamp power Note The lamp must be switched on before operating the computer 24 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Defining an acquisition protocol 4 3 5 Switch on any printer 6 Double click on the icon for the IN Cell Analyzer 1000 Instrument software The IN Cell Analyzer 1000 software will then start 4 3 Defining an acquisition protocol 1 Torun the Acquisition Protocol Wizard open Application gt Acquisition Protocol Manager 2 When the Acquisition Protocol Manager dialog box appears press New Protocol name E Edit _ Mew Import Export Delete Protocol name hg Available in curent setup es Microscopy type Fluorescence Matical_384_0 17mm plate CoolSnap HO Objective Nikon 4s Pland po Fields per well 1 Auto focus search Search 1 0um accuracy 0 4um Combining 2 at 50 00prn Direction Horizontal Snake Number of active wells 384 Time series Off Expected running time Off Cancel Help Figure 4 3 Acquire protocol manager dialog box You will then be asked whether you want to Use currently selected protocol as a template if one has previously been defined If you select No the Acquisition Protocol Wizard starts with default settings If you select Yes the Acquisition Protocol Wizard s
27. on software IN Cell Analyzer 1000 Operating Instructions 28968377 AA 45 4 Operation 4 6 Displaying an image stack To acquire optical z sectioning images you use the same basic procedure as you would for acquiring wide field images Do one of the following 1 Create a new Acquisition Protocol 2 Modify an existing acquisition protocol for wide field images and edit it 46 Displaying an image stack 1 To view the image stack i e all acquired images press the Image stack and analysis icon The Image Stack and Analysis opaan screen O appEa Image Stack Current image stack DAL AIMGSTACE Hello World a 4 sdce i i Yiew current image stack ea re Open image stack file Cancel Figure 4 35 Image stack e View current image stack Work with the image stack that is already open if any e Open image stack file Load an image stack from disk Image Stack D Apps AL33_S IMGSTACK Hello World A A xdce Cc no images acquired partially acquired acquired v Fe gt Time 7 7 zfi fl A Jal gt IV Autocontrast Selection View thumbnails Done Help Figure 4 36 Image Stack window 46 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Procedures after a run 4 7 2 To view individual images click on the well location select the well location from the Well drop down list and or use the navigation keys If multiple fields were acquired from th
28. ons 13 Segmentation parameters are enabled when the Flexible Number of Fields checkbox is marked on the Acquisition Options dialog Segmentation parameters IN Cell Analyzer 1000 Operating Instructions 28968377 AA 33 4 Operation 4 3 Defining an acquisition protocol are used to set the on line cell counting parameters and should be optimized before acquiring images Acquisition Protocol Wizard Segmentation Parameters Specity segmentation parameters hen a new Nucleus wave is selected pertorm digitization to view the updated image Press Helpo for more information T Force call court Fol Nucleus wave D 360_40x HO460_40M Minimum nucleus area 10d ume Segmentation method Celk found Cancel Dra Heb Figure 4 18 Acquisition Protocol Wizard Segmentation Parameters Phase contrast microscopy is a contrast enhancing optical technique that can be used to produce high contrast images of living cells in their natural state without being killed fixed and stained The High pass filter controls the amount of detail that is filtered out of the image The DIC Settings controls the settings for DIC 34 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Defining an acquisition protocol 4 3 synthetic images Acquistion protocol Wizard Phaze Contrast and DIC Specify phase contrast and DIC parameters Phase Contrast Settings High pass filter 0 100 E foo i Invert phase pola
29. other way than described in the IN Cell Analyzer 1000 and or Auto IN Cell Analyzer 1000 manuals WARNING The IN Cell Analyzer 1000 uses extremely high voltage Ensure that all power has been shut off prior to opening the lamp housing WARNING Make sure the rear power connector is always accessible WARNING Use caution when using solutions near the instrument If any liquid is spilled on or around the instrument unplug the instrument immediately and wipe it up If there is any possibility that liquid may have leaked into the instrument case contact GE Healthcare immediately Do not plug the instrument into any electrical outlet until the problem is resolved WARNING Using controls making adjustments or performing procedures other than those specified in the IN Cell Analyzer 1000 documentation can result in hazardous exposure to high voltage or moving parts Exposure to these hazards can cause severe personal injury WARNING This instrument is not designed to the Medical Devices Directive 93 42 EEC and should not be used for medical purposes and or in the diagnosis of patients 12 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Safety instructions 2 Safety precautions 2 1 NOTICE The Pifoc element of the IN Cell Analyzer 1000 is extremely sensitive Any undo force or twisting action can damage the Pifoc crystal Adhere to the warnings marked on both the turret and Pifoc element Using flammable liquids
30. pecified by the Acquisition Protocol to capture images automatically with little or no user intervention The IN Cell Analyzer 1000 uses both laser guided hardware and sophisticated software Auto focus algorithms to automatically find the best plane of focus at each well position When acquisition is complete the entire image set is stored to disk as a single image stack Once an Acquisition Protocol has been defined an image stack can be acquired There are four ways to Initiate the image acquisition process IN Cell Analyzer 1000 IN Cell Analyzer 1000 f Start plate manager Figure 4 27 Start acquisition session e Select the Start acquisition session option from the screen that appears when you enter Assay Development mode e Select the Acquisition option from the Mode menu e Select the Start Acquisition Session option from the Application menu e Click the Acquisition wizard icon from the Application Toolbar Performing any of these operations launches the first page of the Start Acquisition Session Wizard IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Acquiring images 4 5 1 Onthe first page of the Start Acquisition Session Wizard select one of the following options Start Acquisition Session xj Acquisition Do you want to continue acquiring an existing image stack or create a new one Create a new image stack file Continue with current image stack file D Apps 4L33_Shippi
31. r Assay Development mode Plate Manager E lt FPlaterslide holder name New Import Export Delete Rename Edit Flate name Greiner uClear Walid Yes Number of wells 36 8 12 Wall shape Found Plate height 14 00 mm Top left well center 14 36 11 24 mrm Spacing 9 00 9 00 rrn well size 6 50 mm Bottom thickness 250 0 i a4 Cancel Help Figure 4 25 Plate Manager window 38 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Preparations before start 4 4 4 Press New to bring up the Create New Plate or Slide dialog box Plate Editor J Enter plate parameters General Well Layout Plate name No Interval l FEE Rows o mm Plate height mm ooe Columns Ties mrm Bottom thickness um Well Parameters m Well Offset Bottom height mm Round C Square E mm Bothorn interace Plastic At ci Well Volume ul mrm 7 Plate name cannot be empty Cancel Help Figure 4 26 Plate Editor window 5 Select Plate and choose whether you want to Use the following as a template if one has previously been defined Click OK to go to the Plate Editor dialog box 6 Run the Acquisition Protocol Wizard from the Applications menu IN Cell Analyzer 1000 Operating Instructions 28968377 AA 39 4 Operation 4 5 Acquiring images 4 5 Acquiring images 40 The IN Cell Analyzer 1000 uses the information s
32. r diagnostic purposes Safety notices These Operating Instructions contain WARNINGS CAUTIONS and NOTICES concerning the use of the product with meanings as defined below WARNING WARNING indicates a hazardous situation which if not avoided could result in death or serious injury It is important not to proceed until all stated conditions are met and clearly understood CAUTION CAUTION indicates a hazardous situation which if not avoided could result in minor or moderate injury It is important not to proceed until all stated conditions are met and clearly understood NOTICE NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment 6 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Introduction 1 Regulatory information 1 2 Notes and tips Note A Note is used to indicate information that is important for trouble free and optimal use of the product Tip A Tip provides information that can improve or optimize your procedures Typographical conventions Software texts and commands are identified by bold italic text A colon is used to separate menu levels e g File Open refers to the Open option in the File menu 1 2 Regulatory information This section lists the directives and standards that are fulfilled by the IN Cell Analyzer 1000 Manufacturing information Requirement Content Name and address of manufacturer GE Healthcare UK Ltd Amersham Place Lit
33. ra are sometimes affected by a phenomenon known as a stuck charge in which brighter parts of an image are not completely cleared during the CCD readout process As a result these bright areas can appear on subsequent images To remove a stuck charge contact the GE Healthcare support team Replacement of Lamp in Light Source The lamp and bulb are not accessible To replace the light source lamp contact your local trained super user or your local GE Healthcare service engineer Operating Computer Please refer to the manual supplied with the operating computer 50 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Troubleshooting 6 How to get help for IN Cell instruments 6 1 6 Troubleshooting This section contains tips and information for troubleshooting the IN Cell Analyzer 1000 Many problems in instrument loading and cleaning show characteristic baseline and titration abnormalities and knowing which system is affected can greatly expedite the resolution of the issue Some problems can easily be corrected by the average user some may be corrected by the more advanced user and some require the expertise of a GE Healthcare service technician The GE Healthcare service department is happy to provide any advice parts or service that may be necessary 6 1 How to get help for IN Cell instruments Please contact us for any instrument or data analysis questions or issues you may have For contact information for your local offi
34. rity Nikinnimatan method J raremitteditight Reflected light DIC Settings Contrast angle degrees 135 4 Combiner prism dearees 0 Intensity modulation standard Absorphar rate AO 10 I Enable automatic image update lt lt Back Nestes Finish Figure 4 19 Acquisition Protocol Wizard Phase Contrast and DIC 14 Image Combination creates a single focused image from a set of images acquired at different focal planes Enter the size of the Z axis step in the entry field dequisitun Protocol Wizard Image Combination Specify image combination parameters Press Help for more information i Use image combination Number of planes 2 Define combining ENE E Auto Z stepping 50 pm Cancel Finish Help Figure 4 20 Acquisition Protocol Wizard Image Combination 15 Software Auto focus Search feature uses sophisticated software algorithms to automatically find the Z axis position with the sharpest focus This software based IN Cell Analyzer 1000 Operating Instructions 28968377 AA 55 4 Operation 4 3 Defining an acquisition protocol Auto focus feature can be used instead of or in conjunction with the laser guided Hardware Auto focus HWAF system sequisitgy Protocol Wizard Software Auto Focus Search Specify software auto focus parameters Press Help for more information M Use software auto focus search Search fioa um from current position Initial step fio pm
35. service engineer The lamp housing is shown below Bel Figure 1 3 Lamp housing inside enclosure IN Cell Analyzer 1000 Operating Instructions 28968377 AA Safety instructions 2 Safety precautions 2 1 2 Safety instructions This chapter describes safety compliance safety labels general safety precautions emergency procedures power failure and decommissioning of IN Cell Analyzer 1000 2 1 Safety precautions Introduction Before installing operating of maintaining the system you must be aware of the hazards described in the user documentation Follow the instructions provided to avoid personal injury or damage to the equipment The safety precautions in this section are grouped into the following categories General precautions Flammable liquids Personal protection Installing and moving the instrument System operation Maintenance IN Cell Analyzer 1000 Operating Instructions 28968377 AA 11 2 Safety instructions 2 1 Safety precautions General precautions WARNING Provide proper electrical power to the instrument This should be 100 240 Volt 50 60 Hertz alternating current with a Ground Fault Circuit Interrupter GFCI Some power strips such as the one provided by GE Healthcare with your instrument contain a GFCI All power plugs and cords should be 3 prong grounded cables or outlets WARNING In case of fire unplug the instrument WARNING Do not operate the IN Cell Analyzer 1000 in any
36. t pm M Linked Linked i oo o E 5 s e2060 os A al m4 e260 z seoa i m E 5 Fused image ig nable fused image Define Auto offset a Enable exposure and HWAF offset update Cancel lt lt Back Nexb gt Finish Help Figure 4 13 Acquisition Protocol Wizard Filter Selection and Display 50 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Defining an acquisition protocol 4 3 10 Time Series page is used to specify the time intervals between acquisitions for a kinetic experiment If a liquid handling procedure is to be defined it will first be necessary to add a time series In doing so sufficient time intervals must be left between image acquisitions to enable the liquid handling procedure to be completed There are two types of time series available termed Spit and Stare and Look Walk Look modes Time Series Speci parameters for time senes acquisition Acquire time series 0 000 Time units Figure 4 14 Acquisition Protocol Wizard Time Series If the Liquid Handling Module is fitted your IN Cell Analyzer 1000 Instrument contains a liquid handling system for dispensing or aspirating a reagent from 96 IN Cell Analyzer 1000 Operating Instructions 28968377 AA 31 4 Operation 4 3 Defining an acquisition protocol and 384 well plates and for cleaning the system using a wash station Acquisition Protocol Wizard Meazue Figure 4 15 Acquisit
37. tarts with the parameters of the currently selected protocol Enter a name for the protocol Type a name into the entry field All existing protocol IN Cell Analyzer 1000 Operating Instructions 28968377 AA 25 4 Operation 4 3 Defining an acquisition protocol names are listed in the dropdown list lediisitivn Myutuegh Wizard Protocol Name Specify a name for the new protocol and enter a description Protocol name X Description Cancel Figure 4 4 Acquisition Protocol Wizard Protocol Name 3 Specify a password to associate with the protocol Although you will be able to run the Acquisition Protocol without a password the assigned password must be entered in order to modify it slequisitivn Mrutueug Wizandi Protocol Password Specify and verify a password which will be required to modify the protocol Password Verify password Cancel lt lt Back Finish Figure 4 5 Acquisition Protocol Wizard Protocol Password 26 IN Cell Analyzer 1000 Operating Instructions 28968377 AA Operation 4 Defining an acquisition protocol 4 3 4 Specify which microscope objective to use during image acquisition All installed objectives are listed along with a summary of their optical parameters Objective Objective name Objective name Magnification Numerical aperture Refractive index Focal length microns Objective height mrm Camera type Camera width Camera height _
38. tle Chalfont Buckinghamshire HP7 9NA UK Name and ID of notified body INTERTEK SEMKO AB NB 0413 Place and date of declaration Uppsala Sweden Oct 2009 Identity of person authorized to sign See EC Declaration of Conformity Declaration of Conformity CE Conformity Directive Title 2006 42 EC Machinery Directive MD 2006 95 EC Low Voltage Directive LVD 2004 108 EC ElectroMagnetic Compatibility EMC Directive IN Cell Analyzer 1000 Operating Instructions 28968377 AA 7 1 Introduction 1 2 Regulatory information International standards Standard Description EN 61010 1 Safety requirements for electrical IEC 61010 1 equipment for measurement CAN CSA C22 2 control and laboratory use no 61010 1 EN 61326 1 EMC emissions and immunity Harmonized with CISPR Group 1 requirements for measurement 2004 108 EC Class A control and laboratory use EN ISO 12100 1 Safety of machinery Basic Harmonized with 12100 2 concepts general principles and 2006 42 EC design EN ISO 14121 1 Safety of machinery Principles of Harmonized with 14121 2 risk assessment 2006 42 EC CE marking The CE marking and the corresponding Declaration of Conformity is valid for the instrument when it is e used asa stand alone unit or e connected to other CE marked instruments or e connected to other products recommended or described in the user documentation and e used inthe same state as it was delivered from GE Healthc
39. use of a personal computer running Microsoft Windows in the version provided with your product e You should be acquainted with the use of general laboratory equipment and with handling of biological materials e You must read the Safety Instructions in Chapter 2 of these Operating Instructions e The system should be installed according to the instructions in Chapter 3 of these Operating Instructions e You must read and understand these Operating Instructions In this chapter This chapter contains important user information and a general description of the IN Cell Analyzer 1000 and its intended use IN Cell Analyzer 1000 Operating Instructions 28968377 AA 5 1 Introduction 1 1 Important user information 1 1 Important user information Read this before using the IN Cell Analyzer 1000 i All users must read the Safety Instructions in Chapter 2 of these Operating Instructions before installing using or maintaining the system Do not operate the IN Cell Analyzer 1000 in any other way than described in the user documentation If you do you may be exposed to hazards that can lead to personal injury and you may cause damage to the equipment Intended use The IN Cell Analyzer 1000 is an Isothermal Titration Calorimeter system designed for bio molecular interaction studies in research applications The IN Cell Analyzer 1000 system is intended for research use only and shall not be used in any clinical procedures or fo
40. wing message appears Special skills and knowledge are needed to properly calibrate OZ module Do you want to continue Click Yes The Calibrate Optical Z sectioning Module dialog box appears Calibrate Optical sectioning Module OZ Module x Stage Position pr Operations xo 30699 2 Dock Plate Z 23365 Y 3r 00 0 Rough Focus Mave to 2 Move ta xy Settings Bey Step Z Step al al Tools al a l Self Test M z pa Benchmark Current S N 339 Minimum SA 50 Amplitude level agi Status Oz Module last calibration 2006 02 27 12 12 12 Carcel Help Figure 4 34 Calibrate optional Z sectioning module dialog box Adjust the position of the slide so that the light beam from the objective is on the fluorescent part of the slide Under XY Step click the up and down arrows to move the slide in the Y direction Click the left and right arrows to move the slide in the X direction Click Rough Focus Rough Focus finds the approximate bottom of the calibration slide using the Hardware Auto focus HWAF feature Rough Focus brings the bottom of the slide close to the focus point to within 200 um Under Z Step click the up and down arrows to fine tune the focus of the grid lines in the Image View Area Note that the image gets brighter as it gets closer to being in focus 4 Check optical z sectioning movement control 5 Achieve maximum signal noise S N ratio 6 Run the calibrati
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