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ORCA Bioreactor Manual - Harvard Apparatus Regenerative

1. l6 Y barb 721587 yo e 30407 31 n Y la Y barb 30350 JD yd Barb 28 UNF 30407 51 lj AE Bubble Trap 4 30450 AH barb s Right angle 30514 P 4 8 AQ Valve 1 x Diastolic after load pressure 880427 An l4 barb 2 4 NPT 30494 All 1 8 pipe NPT Adapter 30325 AJ Female Luer 1 4 28 30472 All Valve 1 2 Diastolic after load pressure 880426 W Swabble Port 880434 ABD Pulsatile Pump head XXXXX AR Reducing Connector barb 4 Barb 30371 21 June 19 2013 Revision 1 2 PharMed Tubing Reference Guide The following is intended to be a quick reference guide for deciding what size of PharMed tubing is currently being used As previously mentioned it is recommended that tubes of the sizes that your lab commonly uses are glued to a plastic clip board allowing for even simpler comparison PharMed Tubing 13 14 16 25 17 18 Ho2 Front View Doses Tubing size 5 32 4 0mm OD 0 03 0 8mm ID 3 16 4 8mm OD 0 06 1 6mm ID 1 4 6 4mm OD 0 12 3 1mm ID 5 16 7 9mm OD 0 19 4 8mm ID 3 8 9 5mm OD 0 25 6 4mm ID 7 16 11 1mm OD 0 31 7 9mm ID 3 4 19 1mm OD 0 5 12 7mm ID Used for connections not for use in the peristaltic pump Lateral View 29988 1 Note When feeding the tubes through a peristaltic pump it is recommended that you set a convention for flow dir
2. 5 5 pH Sensor Calibration When the pH reading on the DSP4000 is not accurate a 1 point calibration can be performed on the DSP4000 A calibration buffer of a value near the pKa of the sensor is used For a 5 5 10 pH sensor the pKa or midpoint of the response curve falls at 8 2 therefore we choose pH 8 as the preferred calibration buffer for this sensor When complete put the DSP4000 into RUN MODE and collect data into LabChart Switch to pH 6 or other lower pH solution that falls below your optimal area of interest Calibration Solution for 10 minutes or until the reading has fully stabilized and continue to collect data Once done stop data recording and highlight the full section of data that includes both the high and low calibration points Right click on the range in the channel and choose Units Conversion from the drop down menu and use the values that are displayed on the DSP4000 for the 2 point calibration in LabChart 6 7 June 19 2013 Revision 1 2 6 Viewing Data on DSP4000 Run Mode will by default show you the channel measurement data for the sensor plugged into that channel The AUX button changes the display to give current readings of other channel specific Information If the AUX button is pressed it scrolls the screen through displays of temperature pressure AGC Value and Channel Measurements 6 1 Channel Readings O2 CO2 pH Parameters These are displayed as in the image to the left The channel desig
3. HARVARD APPARATUS REGENERATIVE TECHNOLOGY Harvard Apparatus Regenerative Technology 84 October Hill Drive Holliston MA 01746 USA www harvardapparatusregen com 508 893 8999 regenmedicine harvardapparatus com Contents Contents Disclaimer Symbols Overview Chapter 1 Introduction 1 1 Warning amp Caution Statements 1 2 Intended Use Chapter 2 Getting Started 2 1 Safety amp Facility Requirements 2 2 General Equipment Components Chapter 3 Assembly Instructions 3 1 Identifying Components 3 2 Assembling Components 1 Key for Flow Path Setup ii PharMed Tubing Size Reference 3 3 Chamber Specific Flow Path Setup i Large Animal Organ System 14 Chamber ii Large Animal Organ System 10 Chamber iii Small Animal Organ System 5 5 Chamber Chapter 4 Operating Instructions 4 1 Sterilization 4 2 Reservoir Bottle Configurations 4 3 Software 4 4 Heater Temperature Control amp Pressure Channel Calibration 4 5 mage Capture System Chapter 5 Care amp Maintenance Ordering I nformation Replacement Part Numbers Appendix A Polestar Instructions Appendix B Frequently Asked Questions 12 14 20 22 23 23 24 29 32 33 36 49 52 53 54 55 14 June 19 2013 Revision 1 2 Disclaimer Use of the ORCA Bioreactor should be conducted by a trained and manufacturer qualified representative Harvard Apparatus Regenerative Technology does not warrant unauthorized use of this pro
4. Pumps Pumps are selected according to the organ and protocol requirements The ORCA controller along with your pumps of choice address the needs of the decellularization and recellularization processes of various organs Several types of pumps may be controlled including peristaltic pulsatile and syringe A maximum of four pumps are used simultaneously on a system The ORCA controller may be configured to control either four peristaltic pumps or three peristaltic pump heads and a pulsatile blood pump Both configurations typically include two single channel dual head peristaltic pumps A 4 pump may be added to the four unit controller to serve as a spare unit Pump head configurations may be altered at any time to meet the most demanding protocols Selection of the pulsatile pump normally occurs when pulsatile flow rates in excess of 2L min are required This selection must be made at the time of purchase If the four peristaltic pump controller is chosen the fourth pump may be added at any time A fourth peristaltic pump adds flexibility to accommodate elaborate protocols but is not required and can be added at a later time These configurations can be used to accommodate protocols that require a perfusion pump ventilation pump and a pump that will provide flow to the oxygenation system and a gas monitoring system Exterior medium feed and waste bags can also be accessed using channels on the double head double channel pump 9 J
5. StartMode Ramp Start After Time None Time 45 June 19 2013 Revision 1 2 Repeat Mode Repeat Mode Please select the Start Mode vou wish Four modes of repeat are allowed RepeatMode J No Repeat e N ore D e at Repeat Till User Stop Repeat Number of Times Repeat Increase n Times e Repeat until e Repeat of times e Repeat increase N OK Cancel No Repeat mode the step does not repeat Please select the Start Mode you wish RepeatMode In Repeat Until User Stop Mode the step will repeat until pem uon 3 the user stops the program manually The Flip Direction switch allows the flow to be reversed In this mode the step will be repeated for a fixed number of times The Count option allows you to set the number of iterations you want there is no limit RepeatMode The Flip Direction switch up allows the flow to be reversed ade on every other cycle le 10 repeats with flip direction m switch up would result in 1 2 5 the cycles in the forward direction and 1 2 5 in the reverse direction Switch down and all cycles in the same direction In Repeat Increase Mode the speed of the pump increases with every subsequent iteration Please select the Start Mode you wish RepeatMode Count allows you to set the number of iterations Repeat Increase n Keep Pump Running Count Change by allows you to set how much to increase the o speed of the pu
6. Flow Direction aa Changes the direction that the pump rotates a right arrow indicates clockwise Create Step Use this to create a Step See Step Creation for additional info Select Step Use this to select a Step See Step Creation for additional info e Green Light Glows when pump is on vig Step Mode See Step Mode for additional info Start Mode 9 See Start Mode for additional info sme Stop Mode Le See Stop Mode for additional info Repeat Mode See Repeat Mode for additional info Pump Selector Choose from peristaltic syringe pulsed blood or ventilator modes 39 June 19 2013 Revision 1 2 Step Creation Modification In these windows you are able to create a new step or update an existing one The program asks for the user to input a step name and description for each new step or updated step You can see the step in graphical form using the Step Viewer Selector HARVARD APPARATUS Step Viewer Selector Step Mame rar 10 mums Do you wish to create a new step with this configuration or up date we hn the selected step If you select up date any protocols using this step Gp Constant Rate al ca UR vp will be up date also Best practice would be to create new unless you are sure of all usages of this step e Start Immediately Oo Stap on Time Target Step Name O Rate 1001 Run5 165 0 Step Description Start Immeditly Run a Constant Rate 100ml min Infuse Dire
7. June 19 2013 Revision 1 2 Troubleshooting Guide The DSP4000 does Insufficient or too much Disconnect power from the not display power is being applied to DSP4000 meas ETENE or the DSP4000 Verify that the voltage is 24 VDC dii and the current is at least 1 5 Amps Re connect the power The DSP4000 The USB KEY is not Re seat the USB KEY powers up with a recognized Press ENTER USB error The DSP4000 does The AGC for at least one Verify all optical connections not go into Run channel has reached 4000 3 Chil tha ACC Ee euch mode or kicks out the maximum value shortly after because startup channel Replace sensing elements as There is a poor optical needed The Status LED is connection red e sensor element is at the end of its life The Status LED is The AGC for at least one 1 Check the AGC for each yellow channel has reached 3000 channel because 2 Clean the optical surfaces using Dirt has accumulated on Scotch tape as dust and dirt an the optical surface particles may result in signal The sensing element is loss near the end of its 3 Replace sensing elements as useful life needed If none of the above apply and you are seeing AGC Limit please check to make sure that the sensor chemistry is as close to the LED fiber optic as possible If using a glass flow cell check to make sure that it is seated in the cell mount such that the chemistry is between the light and the p
8. Ret Bue 2 7 Period geeet Peale Pume DPM ecu Select Pultalte 58 eu Select 3 Systole mal Select Erde Bool Select Saving Files Be certain to create an experiment folder in my documents by naming the experiment see below Experiment Name Once you save after naming the experiment a folder will be available in the computers My Documents Folder and can be copied onto a portable drive and taken to another PC for evaluation zc X HARVARD APPARATUS 3 6 2013 10 28 43 AM nt Name Experiment Name Status 3 1 Experiment Time Data Start Tme Total Fansed Time 600 amp ep iii ve 100 00 mn y 2 sate Cycle Total Run Tine 2 84 lt T 20 29 Me xc Teme NaN Running Time Press NaN avg Press 280 avg 44 o F avg Rate 0 00 Rate 0 00 eeen Rane 6 05 inn 1 05 Avg Paused Time tecond Temperatures Fressue lowReie lCherts Puree eters Pump 3 5 Pump Charts Ansiog Inputs Logong Rate emp 1 Experiment Log 2 8 3 6 Average PP 4 5 Tre Toncte Pure 58 56 54 52 5 4 AQ 0 36 36 34 32 00 28 26 24 22 2 47 June 19 2013 Revision 1 2 Reading Data Files T GallerySystems In order to read data logged as an Excel file the computer that you are u
9. NPT 1 4 NPT Nuts 1 4 Barb 1 8 NPT 1 4 NPT Nut 34 June 19 2013 Revision 1 2 Luer lock utility port in Return from chamber inch out reservoir to left atrium V aorta trachea outlet Luer lock utility port out chamber or right atrium aorta trachea inlet Temperature probe Manifold allowing multiple feeds or return lines may be created as necessary Add the number of Slip Luer T fittings as required Polycarbonate products are autoclavable They must be thoroughly rinsed before autoclaving because detergent residues cause crazing and spotting Autoclaving cycles should be limited to 20 minutes at 121 C PC shows some loss of mechanical strength after repeated autoclaving and therefore may not function well under high stress applications such as centrifugation Inlet Outlet Adaptor T Fitting 722749 Cap 721449 Clear 1 722734 A Note By convention green caps will indicate inlets to the reservoir while the red caps will indicate outlets Red Nut on outlet 722731 Green Nut on inlet 722732 35 June 19 2013 Revision 1 2 ORCA Controller Software Operation Instructions The operation software allows for control of flow rate and gives temperature pressure and flow rate readouts over time The user can control a maximum of four pumps at once both pulsatile and peristaltic pumps can be utilized IMPORTANT Note Before use of the software be sure to turn
10. Trap Ball 14 chamber 1 ball 30609 10 chamber ball 30564 17 June 19 2013 Revision 1 2 11 There are two fittings on the bottom of the chamber that allow filling and draining of the chamber Add Quick Disconnects to facilitate the connections using an appropriate size of tubing from the chamber fitting 12 The level of liquid in the chamber can be determined by cutting the rigid tubing inside the chamber to the desired height 18 June 19 2013 Revision 1 2 13 Add Temperature probe to reservoir 14 Add appropriate size and length of tubing to the reservoir inlets Note It is recommended that commonly used sizes of tubing are glued to a plastic clip board Take a one inch piece and glue laterally and a 1 4 piece glued so that the internal ID of the tubing is easily displayed This is useful because otherwise it is sometimes difficult to tell the correct size tubing needed at a glance particularly when differentiating between PharMed 17 and 18 See next page for a reference guide 15 Seal the 1 2 inch Barb Fittings that are not in use with caps 30603 This is to maintain the system s sterility 19 June 19 2013 Revision 1 2 Tubing Pathway Setup Key Head Config Typical Function Perfusion into organ for Perfusion pathway into organ Two Single Channel smooth high flow rate Heads Combined by a Y Empty chamber to Oxygenation Polestar pathway reservoi
11. Tubing Size and or add on Calibration Factor selected The calibration factor can also be adjusted Head Configuration Tubing Size Single Head L S 17 Head Configuration Select from single double triple NE A Max 840 ml mi or quadruple head configurations Note The max speed of the motor is 300rpm M Tubing Size Select the PharMed tubing size that you wish to use See table below for a description of the avallable tubing sizes and their corresponding flow constraints When you have selected a particular tubing size the minimum and maximum flow rates for the selected tubing will automatically be displayed below Pump Calibration Pump 3 CAUTION The user must kn OW the tu bing SIZE Please select Head Configuration Tubing Size and or add on Calibration Factor Pump Calibration being utilized for each pump head and verify that the correct tubing size is selected The pump cannot verify this for the user Please select Head Configuration Tubing Size and or add on Calibration Factor Head Configuration Tubing Size Calibration Factor Single Head L S 18 1 Calibration Factor The number set for this parameter m is the inverse of the variance of the liquid delivered The Mes 1140 mi calibration factor is selected based on the deviation from OK Cancel optimal flow rate o The user may wish to run their perfusate at a specified rate into a graduated cylinder in order to test the actual vo
12. make for easier identification later Connect the pressure transducer cable to the ORCA controller 9 When the system is to be used the connectors labeled 1 4 will be connected to the pressure transducers see Operations section for calibration procedure Note Make sure to clean transducers thoroughly before use Pressure transducers are NOT autoclavable Some media types have proteins and other materials that can nonspecifically bind to the pressure sensor and adversely affect the pressure readings One technique used to minimize this effect is to inject 3 mL of sterile water through the swabble port This provides an insulating fluid between the transducer and the medium without a negative effect on the pressure readings Note While not required it is a good idea to line up the Harvard Apparatus logo sticker with the front of the chamber before screwing on the chamber lid 16 June 19 2013 Revision 1 2 10 Place ball on bubble trap base then place bubble trap head on base The actual location of the bubble trap s may vary according to the system being used The location of the bubble trap and number required as well as the right angle fittings may vary depending on the protocol being implemented Note In order to install the right angle fittings the bottom fitting must first be disassembled in order to fit through the Lid hole Swabble ports 880434 Bubble Trap Head 14 chamber 30295 410 chamber 30442 Bubble
13. match the sensor fiber optic cable and channel with the correct parameter units and calibration data that were used during testing Instructions on how to do this are described in section 2 4 of the DSP4000 Optical Process Monitor User s Manual You can change the assignments later by loading each sensor s calibration file to a different channel as discussed in Chapter 3 Managing Configuration Files Removing the USB Key while unit remains under power without powering off the DSP4000 For full instructions please refer to DSP4000 Optical Process Monitor User s Manual section 2 8 60 June 19 2013 Revision 1 2 This is done when you wish to review the log file or update lot calibration information such as when a new sensor is purchased Using this method also closes the current log file such that when measurement resumes you will be recording onto a new log file note this does not work with older USB Keys where the software may freeze 4 2 Connecting RTD Temperature Probes to DSP4000 and Sensors If your RTD Probe s were not purchased with the system and are purchased as an add on they will need to be wired into the DSP4000 Additional information can be found in Section 4 2 of the DSP4000 Optical Process Monitor User s Manual Step 1 Remove the three pronged connector from the end of the RTD Probe using a very small screwdriver and carefully unwind the bare wire terminations and straighten them out Step 2 Power off t
14. past we have seen clinical oxygenators such as Maquet Terumo and Medtronic also used The life span of the oxygenators are dependent on the medium and additives used and well as individual protocols The extraluminal space of the oxygenator is typically supplied with 95 air 5 CO This can be done with a premixed tank of gasses or a gas mixer The OKO system allows you to feed in CO from a tank and air The air can be supplied from a tank from an air compressor or house air If house air is used it is recommended that a moisture trap be added Heater How is temperature regulation achieved The medium reservoir is the primary source of heat and temperature maintenance for the smaller chambers 5 5 and 10 The 14 diameter chamber has a volume of about 38L and as a result requires a wraparound heater to be used in addition to the reservoir It is recommended that the user preheats the medium in an incubator to the required temperature before introducing it to the reservoir The reservoir can be fed from either a bag or a large feed bottle by using one of the dual channel heads of the peristaltic pumps The slow introduction of medium reduces the chance of contamination since the circuit is rarely opened The reservoir heater is limited to a maximum temperature of 39 5 C so that the medium is not detoured If you place medium from a cold room at 4 C the time required to raise the temperature to 37 C will be very lengthy The reservoir bottle
15. the units conversion calibration to your data acquisition system software For units conversion in LabChart follow these steps 1 Connect the Analog Output Cables to the Powerlab 2 Ensure that the Range is set at 10V on the Input Amplifier Set up the channel appropriately in the Channel Settings Window or individually on each channel as per LabChart instructions 3 Press Start and record the upper calibration value that was used for the 1 point calibration Once the reading is stable and you have your sample captured proceed to step 4 Make note of the reading on the DSP4000 unit during this trace recording as this will be the value used in your units conversion 4 Apply the low zero conditions to the flowcell For example for oxygen this may be the upper calibration point of 95 O2 followed by buffer that has been aerated for 30 minutes or which is equilibrated with the aeration gas Continue recording until the stable phase is reached and you have a suitable trace for calibration Make note of the reading on the DSP4000 unit during this trace recording as this will be the value used in your units conversion 5 Highlight the full calibration data trace high and low point for one sensor type at a time and choose Units Conversion from the right click drop down for that channel Follow normal calibration procedure using the values obtained from the DSP for the high and low points 5 3 Oxygen Sensor Calibration It is recommended t
16. turning shaft during long periods of use As an example an intravenous drip bag with water set to between 1 and 5mL per minute to continuously flush the seal wash path could be used Should I sterilize using plasma sterilization or does autoclavation work The materials of construction are Teflon reservoir stainless steel baseplate driveshaft and valves silicone rubber o rings Kynar luer fittings polycarbonate cover and PEEK arbor components All of these materials will withstand steam sterilization One CAUTION however Please disassemble all the valve and driveshaft parts from the Teflon block before steam sterilization The high temperature may cause the machined holes in the Teflon to become deformed by the parts pressed into them Reinstall the valves and driveshaft after the Teflon cools down The Kynar luer fittings have been installed using silicone adhesive to hold them in place and prevent leaks It should not be necessary to remove these fittings from the Teflon because of the sterilization process Can washing be done with reagent These materials of construction will withstand virtually all biological reagents and cold sterilization agents such as Cidex Cidex OPA Mucasol etc ORCA Bioreactor Controller How many channels can the controller support The ORCA 4 controller allows the use of four peristaltic pumps allowing maximal flexibility to support even the most demanding protocols Typically three pump
17. 3 Revision 1 2 Communication cable Power Cable 511301 US 5113010 Euro see parts list for other countries ORCA Controller 880404 Temperature Probe 551492 527560 Heater 880405 110V 880399 220V Reservoir Types 880406 4L 880421 500mL 880436 1L 880437 2L 13 June 19 2013 Revision 1 2 3 2 Assembling Components 1 Place power cord into controller and electric source 2 Connect laptop power cord to the laptop and electric source 3 Connect communication cord to Laptop and ORCA Controller Cable color may vary 4 Connect pump communication cord to the back of the pump and the ORCA Controller It may be easier to identify the pumps if a label is attached to the drive June 19 2013 Revision 1 2 5 Identify any extra pump heads and mounting screws that have been ordered The se may not be immediately needed for the system but should be kept in a safe place Pump heads either single or dual channel Pump head mounting screws mounting hardware for either 2 3 or 4 pump heads 6 Connect heater power cord to the ORCA controller Add liner to heater and wrap it over the top of the heater 7 Attach the temperature probe to the appropriate connector 1 4 Up to 3 can be used with an ORCA 3 1 controller system with a pulsatile blood pump 15 June 19 2013 Revision 1 2 8 Mark the cables on the connector with the number of the channel that they represent This will
18. 9 2013 Revision 1 2 Table Of Contents 1 Introduction Manufacturer s details 2 Safety Note 3 General Description Application 3 1 Measurement Specifications 4 Initial Setup of 5 4000 Unit Quick start Guide 4 1 Installing the USB onto Computer 4 2 Connecting RTD Probes to Sensors 4 3 Integrating Sensors into System Setup 4 4 Connecting DSP4000 to Data Acquisition System 5 Calibrating Sensors General 5 1 Set Barometric Pressure 5 2 Units Conversion Data Acquisition 2 Point Calibration 5 3 Oxygen Sensor Calibration 5 4 Carbon Dioxide Sensor Calibration 5 5 pH Sensor Calibration 6 Viewing Data on DSP4000 6 1 Channel Readings O2 CO2 pH Parameters 6 2 Channel Readings Temperature and Pressure Readings 6 3 Channel Readings ACG Readings 7 Changing expired sensors 7 1 Updating Lot Number for New Sensor Calibration 8 Cleaning and Maintenance 9 Troubleshooting Guide Ordering Information 56 June 19 2013 Revision 1 2 1 Introduction Supplier s detail These Operating Instructions describe the function and use of the DSP4000 Optical Process Monitor This manual is to be considered part of the system for its proper use and operation and should be kept close to the system at all times The information provided on the DSP4000 has to be read in conjunction with the remaining system documentation for the application to which it is being put to use All the information contained in these instructions has been drawn up after ca
19. Apparatus Regenerative Technology partners with leading global scientists to provide specialized solutions The company Is uniquely positioned to develop advanced instrumentation to accelerate regenerative medicine tissue engineering and cell therapy experimentation From the beginning in 1901 Harvard Bioscience companies worked closely with leading global researchers to produce products with the highest levels of performance quality and support necessary for the new challenges of your life science research We look forward to working with you to develop new tools to assist you in solving the new challenges of regenerative medicine from the lab bench to the patient There are thousands of publications in regenerative medicine to stem cell research utilizing Harvard Apparatus products but we are now introducing some newly developed products one for regenerative organ generation and one for small volume cell delivery into organs These products will serve researchers and physicians to accelerate both research and utilization of that research in patients Harvard Apparatus Regenerative Technology provides e Physiological bio sensors and transducers to measure biological variables to better understand animal to cell physiology mechanically chemically electronically e Physiological data acquisition systems with advanced electronics physiological data manipulation algorithms to measure monitor control and understand complex physiological eve
20. H Scale read for the channel from arameter DSPCALIB DAT in the USB KEY Factor 2 d Factor 3 for channels 2 and 4 in 9 0 9 Figure 4 9 ces Lu CO High scale value 0 PPM 5000 PPM read for the channel Wo KE inthe USB KEY in the USB KEY Factor 1 for channel 3 in Figure 4 9 PPB Dissolved PPB 5000 PPB Pun Level PPM Gas Phase OPPM 5000 PPM 180 180 PPB Dissolved 0 PPB 5000 PPB i Level PPM Gas Phase 0 PPM 5000 PPM Dg d8 9 June 19 2013 Revision 1 2 The output range of the analog output is as follows e Minimum measurement 4mA is 2V output e Maximum measurement 20mA is 9 9V 5 Calibrating Sensors General Calibration gas and solution The DSP4000 typically only requires a 1 point calibration at a value near the range in which you will be measuring e DH measurements use a buffer standard of 8 0 e When a 95 02 5 CO2 gas mix is the gas used for aeration and pH maintenance of your perfusion buffer this gas can be used for the 1 point calibrations of both Oxygen and Carbon Dioxide gas saturate a non bicarbonate buffered solution your perfusion system requires buffers to be deoxygenated such as In lung applications or use buffer equilibrated with room air then air saturated solution non bicarbonate buffered should be used for calibration In these situations it is common to use some 5 CO2 gas mix for the 1 point calibration of the CO2 sensor such a
21. SING A di S OUT DISPOSABLE pH CO2 O2 SENSING ELEMENT Puncture probe 1 Remove the puncture probe from the fiber optic cable This sensor is purchased as in the catalog as it has no replacement portions as it is a disposable item 2 Update the configuration file on the USB KEY as described in Loading the File onto the USB KEY Section 3 4 of the DSP4000 Optical Process Monitor User s Manual BioProbe 1 Unscrew the tip from the BioProbe by twisting it counter clockwise CAUTION The sensing element is designed to seal when finger tight Never use pliers or any other tool to loosen or tighten the sensing element as the tools can damage the sensing element and probe body 2 Screw the replacement element onto the end of the probe Calibration information for the sensing element is contained in a calibration file that includes the sensor type and the calibration factors for all elements in the same manufacturing lot You can download the calibration file from the Support area on the Polestar Technologies web site 3 Update the configuration file on the USB KEY as described in Loading the File onto the USB KEY Section 3 4 of the DSP4000 Optical Process Monitor User s Manual 7 1 Updating Lot Number for New Sensor Calibration See Chapter 3 Managing Configuration Files of the DSP4000 Optical Process Monitor User s Manual for using the Configuration Utility to assign new lot numbers to the measu
22. alibration process Failure to use a flowing calibration solution can affect the calibration results flow through a flow through sensor or use a magnetic stirrer If calibrating with gas the gas should flow through the sensor for at least 5 minutes Application Note If you wish to scale the OUTPUT of the DSP4000 for higher resolution within a smaller range of the total range of the sensor use the DSP Configuration Utility see section 4 6 Scaling Values in the Calibration File of the DSP4000 Optical Process Monitor User s Manual Temperature Influence on CO2 and O2 Measurements For accurate calibrations and measurements the temperature needs to be known and actively compensated You should have the temperature either manually set at the temperature at which the calibration is taking place you can put a small temperature probe into the line to measure or you can use an RTD probe that is wired into the DSP4000 unit The RTD probe is the preferred method as you will have continuous temperature compensation and therefore will not need to change any temperature settings on the DSP4000 between calibration and measurement The DSP4000 can be configured such that a single RTD probe can provide the continuous temperature source for all channels This is appropriate for a sensor like the Tri fold Flow Cell If sensors are located in different areas then multiple RTD probes can be used A Channel is instructed to look at either a Common So
23. atory s individual protocols Small Animal 3 4 tube Pulsatile for trachea 1 8 tube Polestar transducer 3 8 rigid tube PA on lung or pressure control for valve peristaltic 3 8 tube PA on lung or pressure con trol for valve peristaltic 1 8 rigid tube Polestar transducer 1 4 rigid tube chamber circulation 1 4 tube chamber circulation 1 4 rigid tube oxygenator circulation 1 4 tube oxygenator circulation 1 8 thermocouple 1 2 tube Pulsatile for trachea Note This diagram depicts typical sizing amp usage for each of the reservoir bottle ports in a small animal system Be sure to follow your laboratory s individual protocols 33 1 8 tube 1 8 rigid tube Polestar transducer 1 4 rigid tube PA on lung or pressure control for valve peristaltic 1 4 rigid tube PA on lung or pressure control for valve peristaltic 1 8 tube Polestar transducer 1 4 rigid tube chamber circulation 1 4 rigid tube chamber circulation 1 4 rigid tube oxygenator circulation 1 4 rigid tube oxygenator circulation 1 8 thermocouple 1 8 rigid tube June 19 2013 Revision 1 2 Cap mounting Plastic Bottle Large Animal Modified 1 2 Barb 3 8 NPT Modified 1 8 tube 1 8 NPT Modified 1 4 Barb Luer Lok 1 4 1 4 NPT 28 1 8 barb Caps are available to seal the 1 2 inch Barb Fittings gls mat 1 2 Barb 1 4 NPT 3 8 NPT Nuts 3 8 Barb 1 8
24. c bs ra 22 i Pd E A T Ja PharMed L S 16 1 3 PharMed L S 17 3 3 Y Barb Ne Ne X YBb 29 June 19 2013 Revision 1 2 di n Pharmed 25 g Pharmed June 19 2013 Revision 1 2 5 5 Chamber Lid continued Fill from reservoir Reservoir Pressure Left Atrium ventric lar pressure Le Left Atrium w ntricular 31 Right Atrium ventricular pressure Extra probe ports Temperature probe x 2 JA Right Atrium ventricular M Bubble trap empty port Rod mounti ng for manipulator holder June 19 2013 Revision 1 2 Chapter 4 Operating Instructions Prior to Starting Assure that all the requirements identified in Chapter 2 have been successfully met Personnel using the ORCA Bioreactor should read through this manual in its entirety prior to using the device Failure to follow aseptic techniques and failure to train on all processes and procedures prior to using the bioreactor may result in critical delays contamination and other harmful events 4 1 Sterilization Sterilization can be performed using EtO or by autoclaving It is strongly recommended that all fittings be loosened prior to sterilization procedure Tubing has a tendency to form around the barb during an autoclave cycle and can become loose and a
25. can be fed into the DSP unit an accurate baro metric pressure should be read from a NIST certified barometer and manually entered into the unit on a daily basis Instructions for manual input of pressure are found in section 5 6 1 of the DSP4000 Optical Process Monitor User s Manual Options for continuous barometric pressure compensation are RM Young 61302L Barometer with 4 20mA analog output standard Vaisala BAROCAP Digital Barometer PTB330 Manual Pressure Input You can manually enter a pressure value in units of mmHg or PSI Input values must be absolute pressure that is barometric pressure plus gauge pressure The default value is 14 7 PSI 760 mmHg To use a manually input value for pressure compensation 1 Select Pressure from the Setup menu and press ENTER to display the current setting and then press ENTER to show the pressure input options 2 Select one of the manual input options and press ENTER to show the currently specified input 3 Press the up or down arrow to increase or decrease the setting one unit at a time or hold down the key to rapidly change the value 4 Press ENTER to accept the displayed value and return to the Setup menu OPTICAL PROCESS MONITOR PN t 65 June 19 2013 Revision 1 2 5 2 Units Conversion Data Acquisition 2 Point Calibration After the DSP4000 Optical Process Monitor has undergone the 1 point calibration s for the channel s installed you may proceed with applying
26. chamber and use of clear materials allows for easy visibility of the organ being studied Chamber orientation can be adjusted depending on the decellularization and cellularization procedures Chamber access ports are built in for removal and addition of media Numerous sampling ports are included and the built in windows allow for manual access to an organ Hollow Organ Chambers 880403 The rotating double chamber is specifically designed for cell seeding and culturing both surfaces of a tubular matrix The intraluminal and extraluminal flow paths may be connected or maintained separately The hollow organ chambers allow seeding and culturing of different cells types on either side of the tubular structure providing homogeneity The design allows for enhanced oxygenation and mass transport between the medium and cells Large and small animal versions are available 11 June 19 2013 Revision 1 2 3 1 Identifying Components This is a typical system Actual components will vary according to the system ordered Please check your order and shipping manifest for an exact list of components Pump Cables 5155720 Pump Drive 881000 with appropriate heads The type and number of heads will vary with each system 4 channel controller cable 880366 Transducers 880360 are shipped in sealed sterile packaging Pressure Calibrator 734479 Laptop and power cables will vary depending on the order 12 June 19 201
27. ction Run for 5Seconds o iw Woe o cne 60 25 50 7 5 10 0 125 150 17 5 22 0 22 5 25 0 27 5 30 0 32 5 55 0 345 40 0 42 5 45 0 47 5 500 52 5 55 0 am 1 Tene sec HARVARD APPARATUS Step Viewer Selector Create or Select Step Step Name case ot Runs Would you like to Create a New Step with your present configuation Step Description Start 1 89 357 Run a Constant Rate 150m mn Infuse Drection Run for SSeconds sgh Constant Rate 3 Start Immediotely W Create Step Name Rate 1501 Runs oO Stop om Time Target Select Repeat Increase n x select 165 0 14 0 Start Immeditly Run a Constant Rate 126 0 i5 ml min Infuse Direction Run for 55econds 100 0 t 80 0 60 0 3 0 2 0 ITEM HARVARD APPARATUS step Viewer Selector When the step parameters have been selected a new yellow gear with a green E T TT CrOSS will pop Up WZ Constant Rate 3 start immediately You must press on this gear and save the man step for it to be implemented Otherwise Wat ae nu ce me nae m Desin any changes made will be discarded upon CE leaving the window 1 40 June 19 2013 Revision 1 2 Pump Calibration To access the pump calibration menu click on the image of the pump that you wish to calibrate From this menu the head configuration and the correct tubing size can be Please select Head Configuration
28. ctions in the DSP4000 User s Guide e You need to upgrade the operating system to a new version The DSP4000 will prompt you to upgrade the software when you restart the system see Figure 2 12 e There will be an extended period between uses 59 June 19 2013 Revision 1 2 Refer to Quick Start Guide for connection of Fiber Optic Cables NOTE You must align the small tab on the ST Connector on your sensor with the notch in the Fiber Optic cable or the fitting will not connect Recommended not to re screw lid until your setup is complete and the unit is fully calibrated and ready to use NEVER use a Power Strip to power on and off your DSP Unit This results in a fuzzy power cutoff that interferes with the memory and proper functioning of the unit Aw A POLESTAR OPTIC AI DDMCECe HAITAN Proper methods for powering down the DSP4000 Go to Main Menu and select Turn Off DSP4000 This allows you to put the unit into standby and avoid having to wait for a 60 minute warm up period at the next use you wish for a full power shutoff then after the above step open the DSP4000 and flip the toggle switch to OFF Note Saving the Log file is optional If you have performed a calibration procedure or changed any other setting or recorded measurement data onto the log file it is advisable to save the log file 4 1 USB Key t is important to identify which sensor was assigned to each channel in order to
29. date the RTD probe 61 June 19 2013 Revision 1 2 4 3 Integrating Sensors into System Setup The Flow cells in the cell mounts have male and female luer terminations Make sure that you always use a non gas permeable or low gas permeable tubing such as Tygon R 3603 or Tygon S 50 HL Using a Luer Barb connector kit you can fit multiple barb sizes to the luer terminations or connect these to each other end to end for multiple sensor types The Tri fold Flowcell has four threaded holes set in a square configuration 0 20 deep 10 32 Thread M5 Thread to accommodate multiple mounting options It can also be placed on the lab bench or clamped using rod clamp system to the relevant portion of the perfusion system DISPOSABLE pH CO2 O2 SENSING ELEMENT 4 4 Connecting DSP4000 to Data Acquisition System e If you purchased a DAQ DSP4000 Interface Cable this has come preinstalled and configured to work with a standard 0 10V Analog to Digital Data Acquisition System such as a PowerLab Datag NI DAQ iWorx Biopac or other These cables terminate in a Male BNC connector Utilizing DAQ hardware that has analog inputs other than female BNC requires the use of an adapter to your input type or removal of the BNC connector to bare leads e Connect the BNC output cables to the Analog Input BNC connectors on your Data Acquisition System Please connect to the channel on which you wish the particular reading to be recorded Sensor pH p
30. duct Harvard Apparatus does not warrant that the operation of this product will be uninterrupted or error free and makes no claim of warranty or condition Harvard Apparatus reserves the right to change the instructions for use and any related products at any time without any prior notice and is not liable for any damages arising out of any change and or alteration of the contents or product This product is for RESEARCH USE ONLY Copyright 2013 Harvard Apparatus All rights reserved ORCA Bioreactor is a trademark of Harvard Apparatus Harvard Apparatus a division of Harvard Bioscience owns the intellectual property rights to the ORCA Bioreactor This material may not be reproduced displayed modified or distributed without the expressed prior written permission of the copyright holder U S international and foreign patent applications are pending 3 June 19 2013 Revision 1 2 Symbols Used Date of Manufacture Serial Number Catalogue Number Warning This action will have a direct impact on the patient Caution This action will have an impact on the product or operator Manufacturer This device complies with Directive 2006 95 EC relating to electrical equipment designed for use within certain voltage limits this device al so complies with Directive 2004 108 EC relating to electromagnetic compatibility 4 June 19 2013 Revision 1 2 Overview HARVARD APPARATUS REGENERATIVE TECHNOLOGY Harvard
31. e is the maximum rate the pump is allowed Min rate is the minimum rate the pump is allowed Please select the Start Mode you wish PID Setting P Gain is the proportional gain which dictates Run Mode how fast the pump is allowed to change to obtain the Constant Pressure required setting Operation Made PID Setting Gain is the integral value used to adjust the ek seit he NEER Run Mode baseline and fine tune the rate at which the pump is Constant Pressure AvatagaMode i allowed to move to reach the programmed setting Pressure mmHg 1140 Users can also choose between Standard Mode and the PID Settings Average setting ia WE ES Standard Mode uses the raw data of each reading The P Average setting takes a rolling average of the last 40 0 000 readings A reading is typically about 20 milliseconds This setting is often used to smooth out the baseline QmETUEM 42 June 19 2013 Revision 1 2 In Pressure Oscillation Mode the system alternates between using the high rate to obtain the Please select the Start Mode you wish high pressure target and using the low rate to obtain en Madi zi the low pressure target The pressure targets are in units of mmHg and the peres Target ri rates are in ml min High Rate ml min 0 000 In Pulsatile Cycle Mode the program allows for a TUER pulsatile flow to be provided by the peristaltic pumps Please select the Start Mode wou wish Ru
32. ection that you follow for all pumps i e forward flow is always designated as flow from left to right This will help avoid potential confu sion 22 June 19 2013 Revision 1 2 3 3 Chamber Specific Flow Path Setup Large Animal Organ System 14 Chamber Note The following is a diagram representing a configuration for the bioreactor for a arge animal lung system This diagram is not a replacement for your laboratory s protocols be sure to follow your individual experimental procedures Your individual laboratory s setup may differ from what is depicted below G3 32 Oxygenator 1 A3 E d 1 inu ini Gas out Pre Organ 032 po feet A PharMed L S 18 PharMed L S 18 23 June 19 2013 Revision 1 2 Large Animal Organ System 10 Chamber Aorta Pressure 2 Right Atrium Pressure Spare Plug Port Left Atrium ma Pressure 1 Ps N pu di Chamber or Spare Pressure 4 Chamber Vent Organ elevator handle Compression lock screw Right Ventricle Temperature Probe Left Ventricle Pulmonary Ar tery Arm Articulator holder Aorta Feed T Additional Sensor Port 30470 Chamber Quick Connect Fill Chamber Quick Connect Drain 130607 130607 24 June 19 2013 Revision 1 2 Large Animal Organ System 10 Chamber cont 25 June 19 2013 Revision 1 2 Oxygenator A3 Large Anima
33. erfusate media 72 June 19 2013 Revision 1 2 Ordering Information 88 0258 O Replacement Kit sensor glass window amp O Ring 88 0260 pH Replacement Kit sensor glass window amp O Ring 88 0261 CO Replacement Kit sensor glass window amp O Ring 880258 O Replacement sensor 880260 pH Replacement sensor 880261 CO Replacement sensor REPLACEMENT QUARTZ 02 PPB SENSOR INSERT FOR SINGLE QUARTZ 88 0263 FLOWCELL MOUNT 72 8161 BUFFER SOLUTION pH 8 00 500ml 72 8162 BUFFER SOLUTION pH 7 41 500ml 72 8163 BUFFER SOLUTION pH 6 00 500ml 72 8164 BUFFER SOLUTION KIT pH 6 7 41 8 500ml EA 880194 Trifold Connections Kits 1 16 880195 Trifold Connections Kits 1 8 880196 Trifold Connections Kits 1 4 7 UI June 19 2013 Revision 1 2 Appendix B Frequently Asked Questions Hollow Organ Bioreactor Is the retainer clamp missing from my unit The retainer clamp design did not fit as well as it was supposed to and since it did not seem to be necessary we have not been shipping it with the product A new clamp is in the process of being developed to make sure the drive mechanism does not slip and disengage itself How does the seal wash inlet and outlet work for hollow organ systems This passage is designed to take a non salt solution and flush it through a separate space between the two outside o rings on the driveshaft This will prevent any buildup of evaporated salt crystals from forming on the
34. for which the connected sensor s is are configured pH O5 CO The sensors are supplied in a range of configurations to meet the various application specifications of the end users Most frequently researchers who purchase the system from Harvard Apparatus will be utilizing flow through sensors for use in one of the many perfusion systems available however in vivo applications are also common in which a puncture probe may be used General purpose probes can also be used and immersed in any fluid O2 and CO2 sensors can operate with fluid or with gas and therefore a wide variety of applications is possible The DSP4000 can be configured for 1 2 3 or 4 channels for one two or all three of the parameters currently available pH O5 CO A dual channel system can be configured for measurement of 2 of the same parameters requiring only one purchase of the firmware for that measurement or 2 differ ent parameters as long as the firmware for the parameters of interest are both purchased or 3 differ ent parameters with only 2 of the 3 sensors connected at any one time This manual supplement is primarily designed to guide researchers utilizing the system for isolated perfused organ and tissue ap plications or who will be calibrating their sensors in the ranges outlined in section 3 1 of this docu ment 3 1 Measurement Specifications Oxygen Measurement ranges available High 0 500 Air Sat Low 0 20 Air Sat Units of Measurement a
35. g with the Calibration button to calibrate pressure readings The zero is normally set by opening the transducer to air Wait until you have a stable reading one that stays constant for the duration of two flashes of the green light WARNING You must have a pressure measurement device and a way to increase the pressure IF you press the calibration button and do not have a way to measure the actual pressure you will establish a false reading and not be B able to use it typically 100 mmHg Manual Pressure Calibrator 73 4479 49 June 19 2013 Revision 1 2 Chart Readouts Note In any tab you may click on the top and bottom numerical values on the y axis at any time to change the max and min values Click on the actual high number it will light up manually type in the high level you want Repeat this for the low value of the Y axis This is done as if you are changing a text value Also be sure to check the boxes next to the channels that you would like to have plotted Temperatures Pressure Flow Rate Pump 1 Charts Pump 2 Charts Pump 3 Charts Pump 4 Charts Analog Inputs d Temperature 1 Temperature 2 r Temperature 3 n Temperature Temperature c 07 15 14 I l l l l I l l l l I l l l I l l l l I l l l l a t4 1 7 1 1 7 1 ILS WIE IM HR ey Wax G eel a Aa F Ao a An 2 LS I us Time seconds The first tab is for
36. ge M 4 QOO K ek Pulsatile Cycle Press 2 49 mmHg I Avg Press 0 05 mmHg 7 Avg Rate 0 00 ml min Rate 0 00 ml min Pump 3Charts Pump 4Charts Analog Inputs Mel x Pump 2PressureRaw v 3 75 UCL E 3 5 r Je 3 25 Do VAS m LoL ph J2 2 25 2 400 Flow Command CH 1 300 E 200 ed 0 100 200 300 When the trials are running the master controls will appear as above Use the red button to stop all data logging 37 Master Controls Starts all pumps Stops all pumps Incrementally adjusts flow rate of all pumps Adds an Experiment note HARVARD APPARATUS 3 5 2013 10 28 43 AM Experiment Name Experiment Name Status Experiment Time Data Start Time Total Elapsed Time Total Run Time Running Time Paused Time Logging Rate 1second Lg T Experiment Log pa 100 Time Function Pump Value June 19 2013 Revision 1 2 Starting Pump Programs The green icon starts the programs of all active pumps and begins data logging Before data logging begins the user is prompted to either create a new file or append If the create option is chosen but the file name has already be taken the program will issue a warning The disk icon indicates that you are collecting data Experiment Log All data will be cont
37. he AGC level of each channel to identify the channel causing the Status LED alert See AGC Display Section 5 8 of the DSP4000 Optical Process Monitor User s Manual for information on accessing the display from the Setup menu 2 Inspect the BioProbe connections and the optics on the problem channel to determine whether the high AGC is caused by an obstruction or break in the optic path See Inspection Procedures in Section 6 1 1 of the DSP4000 Optical Process Monitor User s Manual 3 Replace the sensing element if the AGC is still high after resolving any problems with the fiber optic path Flow cells glass insert with chemistry painted internally 1 Remove the glass insert by loosening the screw with an allen hex wrench and sliding the flow cell out re 2 Contact Harvard Apparatus or your Harvard Apparatus Appointed Distributor for the correct placement part number for the flow cell insert for your measurement i e O2 PPB O2 PPM O2 Sub PPB CO2 pH 5 5 10 or 4 7 Tri Fold sensor 69 1 Remove the Fiber Optic cable s 2 Using a wrench if unable to remove with fingers unscrew the ST Connector on the channel that needs a replacement sensing disk 3 Contact Harvard Apparatus or your Harvard Apparatus Appointed Distributor for the rect replacement part number for your sensor type e O2 PPB O PPM O Sub PPB 02 pH 5 5 10 or 4 7 June 19 2013 Revision 1 2 x SENSOR HOU
38. he DSP4000 unit by turning it off in the menu and then with the toggle switch inside the case Unplug the unit from the AC Adapter Step 3 Using a small flathead screwdriver loosen the metal clamps for the applicable RTD inputs Step 4 The four cord grips on the bottom panel enable you to bring connecting wires into the DSP4000 enclosure while maintaining a water tight seal To bring the wire into the DSP4000 enclosure e Loosen the locking nut on the cord grip and remove the cylindrical metal plug Figure 4 2 shows a DSP4000 where the left most and right most cord grips have already been used for the power cable and an RS 232 connection respectively The plug has been removed from the grip next to the RS 232 cable Terminal Labels Channel 1 RTD1 Black 1 RTD1 Red 2 RTD1 White 3 Channel 2 RTD2 Black 4 RTD2 Red 5 RTD2 White 6 Channel 3 RTD3 Black 7 RTD3 Red 8 RTD3 White 9 Channel 4 RTD4 Black 10 RTD4 Red 11 RTD4 White 12 A eseesesceceetesaee 4 20 mA Inputs Temperature Pressure 4 20 mA Outputs Mr 55 1 e Store the cylindrical plug in a safe place Step 5 Insert the bare wires as per the above table VAN Clamp down using screwdriver NOTE The white wire is if Power Cable now black the black are interchangeable Connect the threaded RTD probe tip into your sensor The Flowcells and the Tri fold Flowcells both have built in threaded ports to accommo
39. ility is able to provide a clean safe and suitable area for aseptic cell processing It is recommend that all manipulations of the unit once sterile are performed in a biological safety cabinet laminar flow hood Failure to provide a means to conduct aseptic cell processing may result in harmful contamination 7 June 19 2013 Revision 1 2 2 2 Equipment Components 1 Store the ORCA Bioreactor in a cool dry place free from dust and other potential contaminants until ready to use 2 Assure that the ORCA Bioreactor package was received completely and without damage if the package arrived as damaged contact your local technical support group 3 Ensure that all parts of the ORCA Bioreactor package and its peripheral devices are in good condition Do not use damaged parts Overview of Main ORCA Bioreactor Components A Heater Reservoir amp Temperature Control System B Peristaltic Pumps C Organ Chamber D Pulsatile Pump 8 June 19 2013 Revision 1 2 Heater Reservoir The heater is used to maintain temperature throughout the system to emulate physiological conditions The reservoirs hold a volume of 0 25L to 4L of media The reservoir has built in ports for interaction with the oxygenator and the organ chamber as well as a port for a temperature monitoring probe There are different sizes of bottles large and small flow that have different size ports to accommodate a wide variety of tubing and flows
40. inuously entered into the experiment log until the user stops the run Experiment Notes Clicking on the Notepad icon allows the user to enter an experiment note Experiment Note Please enter your Experiment Note below Mote Number 1 Experiment File Name Exists2 Experiment File Name already used Do you wish to Append or Create New Experiment Name Experiment Mame ARVARD APPARATUS 3 5 2013 10 38 20 AM Experiment Name Experiment Name 1 1 Status Running Experiment Time Data StartTime 3 9 2015 10 38 12 AM 00 00 08 00 00 08 00 00 08 00 00 00 Total Elapsed Time Total Run Time Paused Time Logging 826 isecond 1111 3 5 2013 10 38 08 AM Pump OFF 1 3 5 2013 10 38 08 AM 218656 5 52013 10 58 12 AM Set 3 5 2013 10 38 12 AM 3 5 2013 10 38 12AM Pump ON 2 St BP June 19 2013 Revision 1 2 Rate 1501 Runs 150 00 ml min e 100 00 ml min Constant Rate T Constant Rate ec Temp 2284 Temp 39 39 lt Press NaN Press 2 60 mmHg Ava Kate 6 00 ml mi Fate 0 00 ml min 3 Enable Disable Pump This primes a pump for use or deactivates a pump when not in use Start Stop Run Starts or stops the corresponding pump A Flow Rate Control These up down arrows manually adjust the flow rate while the run takes place The flow rate can also be altered by clicking on x xx ml min just below the pump icon and entering a value S
41. l Organ System 10 Chamber cont Note The following is a diagram representing a configuration for the bioreactor for a arge animal lung system This diagram is not a replacement for your laboratory s protocols be sure to follow your individual experimental procedures Your individual laboratory s setup may differ from what is depicted below Pre 26 Organ O2 I 1 Tefombm w 9 89 AR Reducers June 19 2013 Revision 1 2 Large Animal Organ System 10 Heart Chamber Pulmonary artery Right Ventricle In from aorta 4 Out to Reservoir i Out to peristaltic pump 27 June 19 2013 Revision 1 2 1 2 Diastolic after load pressure valve Tighten clockwise to increase pressure 28 June 19 2013 Revision 1 2 Small Animal Organ System 5 5 Chamber Note The following is a diagram representing a configuration for the bioreactor for a small animal lung system This diagram is not a replacement for your laboratory s protocols be sure to follow your individual experimental procedures Your individual laboratory s setup may differ from what is depicted below X i ARX B 2 E g Oxygenator E U2 l Ui NETS in X x X 1 A P U A3 ee f Us t A3 7 Pre Organ O g A d e n te Ju a TL 4 en i Y A 4 e i a
42. llow leaks One possible sterilization procedure would involve assembling the tubing set as it is to be used in the bioreactor and placing it into a sterilization bag to be autoclaved the chamber could then be sterilized in a separate sterilization bag Be sure to follow your laboratory s individual sterilization protocols It is also strongly recommended that PharMed tubing be used throughout the system It has shown to be the most resilient to the intense heat and pressure of the autoclave Tygon tubing is very susceptible to warping after autoclavation If the flow path is autoclaved and has Tygon tubing check all barb connections In many cases Tygon tubing will become soft and may slide off the barb after autoclaving when pressure is applied If the tubing is soft on the barb it is recommended that the portion of the tubing that was on the barb is cut and discarded and that fresh tubing is slid onto the barb Ty wraps may also be used on barb fittings 4 2 Reservoir Bottle Fittings amp Typical Usages Note The reservoir bottle for the ORCA Bioreactor comes in a variety of configurations The various models are all displayed in the next few pages of this manual Please find the model that was included in your laboratory s particular bioreactor package 32 June 19 2013 Revision 1 2 Large Animal Note This diagram depicts a typical usage for each of the reservoir bottle ports in a large animal system Be sure to follow your labor
43. lume delivered If the volume delivered is less than the expected amount the calibration factor can be set at a value greater than one to compensate for this deviation Specifications for Pharmed tubing ne 1 6 Mae 1 6 3 2 4 8 147 6 4 3I 9 5 12 12 7 EE om om om ow ow ow Part Number 25 foot roll 7 6m 720958 Used for connections not for use in the peristaltic pump 41 June 19 2013 Revision 1 2 4 Max Pressure continuous psig bar 25 1 7 25 1 7 15 1 0 10 0 7 10 0 7 0 15 1 0 Operation Mode Operation Mode On this screen Run Mode Flow Rate and Flow Direction can rr hen be controlled y Constant Rate Btandard Mode Flow Direction Constant Pressure Pressure Oscillation Pulsatile Cycle mpeg 570 Run Mode There are four options to choose from e Constant Rate provides a constant perfusion rate e Constant Pressure pump 1 alters its flow to maintain constant pressure Lo o c e Pressure Oscillation adjusts the flow to bounce between two set pressure points e Pulsatile Cycle provides a pulsatile flow cen Standard Mode Flow In Constant Rate mode flow is set by the user and heden maintained at a constant rate Flow direction may be 100 E changed In Constant Pressure mode constant pressure is set by the user and the pump varies the flow to maintain the pressure setting Max rat
44. monitoring the temperatures over time of the channels up to four that you have in use Temperatures Pressure Flow Rate Pump 1 Charts Pump 2 Charts Pump 3 Charts Pump 4 Charts Analog Inputs Pressure 1 a Iv 100 Pressure 2 ES Pressure 3 n 1 Pressure 4 n r Pressure 6 I 1 l 1 1 l A a L5 1 Mh 0 Time seconds This is an example pressure readout for the various pumps flow paths The pressures corresponding to pumps 1 2 and 4 are being displayed Note you do not see all of the traces that you are expecting one or more of the traces could be overlaid In order to remedy this turn off traces sequentially by unchecking the corresponding boxes next to Pressure 1 Pressure 2 etc 50 June 19 2013 Revision 1 2 Temperatures Pressure Flow Rate Pump 1 Charts Pump 2Charts Pump 3 Charts Pump 4Charts Analog Inputs Pumpi je Iv e 230 Pump 2 Iv Pump 3 v Pump 4 et Iv E fen 5 Et LL I i i I i Lib OE 39 30 e o x Time seconds The third tab is for monitoring the flow rates over time of the channels up to four that you have in use Temperatures Pressure Flow Rate Pump iCharts Pump 2 Charts Pump 3Charts Pump 4Charts Analog Inputs Variance Pump 2 Pressure Average Pan Iv zi 100 Pump 2 Pressure Raw unm W 3 75 ba tn UCL yia Pressure mm ha EU 4 in 1 tu LOL n Flo
45. mp for each time the step is repeated Max value Is the maximum flow in ml min this ts limited by the tubing Keep Pump Running allows the pump to continue to run as 46 the final flow rate is reached June 19 2013 Revision 1 2 Notifications This window allows the user to enter c contact information in order to be Notifications User List HARVARD APPARATU notified by email when certain events men occur At present text messaging is not active so entering the phone number has no effect This screen allows you to select specific events for which you will receive notification Notifications System Status HARVARD APPARA 3 5 2013 8 39 47 AM User 66 9 1 V Enable Internval Sewe F Email TextMessage Window Print temp T Pomp Cmimet How Pump i Tenperture Select Pump 2 Temperture Select Pump 3 Tesperture Select Pump 4 Temperture Select Leesten 1 02 Select Pump Freire I Tump 2 Procure Sekel Pump 38 Sheri Pump4Prexase Location 1 12 Seker Pump Liae e Select Fumo Rate 56 2 Fumo dace v detect Pump thae Select testen IMP See Fuso 18625 Tee Select Puro 286 Time Skeet Pume 285 Tine Sect Pump 48866 Tine se 6 85 2 wO2 sect Pump i Dessert Tee Sedert Pump 2DesentTme Gekect Pump descent Time Sert Pump diiesend Time Select Burs Period D meet Pump Bend
46. n i n TUI Fr 1111 j Feu ae U M 1 5 I I l l l l l I l l l l l I 171 a biel Sly Sr Se See rly DS HU ee in Ge SF hg el ten Gel du c Tn I cu mile QC I Time seconds 48 June 19 2013 Revision 1 2 Heater Temperature Control amp Pressure Channel Calibration Heater Temperature Control Heater Temperature Control Access the Temperature Control menu through Tools gt Heater Heater Controls Probe Ta Control Probe to Control allows the user to select between four nicae f different probes labeled TC 1 to TC 4 Heater Control Heater Control can be set to either Enabled or Disabled Disabled SetPoint allows the user to set the temperature in degrees Celsius The Pressure Channel Calibration menu can be accessed through 7ool s gt Calibration gt Pressure Pressure Channel Calibration Please select the pressure channel you wish to calibrate First select the desired pressure channel You can have up to four channels The Filter may be set to either Mean or Raw Actual Value The Actual Value is the instantaneous value for the pressure 2 red d g Cal Value Average Value Average Value is the average reading over a few seconds The Average Cycle Average Cycle serves as an update light to let the user know when the average has been updated Use the Zero button to zero the actual value reading and Cal Value alon
47. n Mode There are five modes of operation Pulsatile Cycle A cer Sta Set Pressure Mode Inverse Mode Set Rate mode Set pressure mode Inverse mode Reverse mode systole 0 000 Reverise Mode m NN In Standard Mode the parameter BPM allows the beats per minute to be set Please select the Start Mode you wish Run Mode Pulsatile Cycle Standard Mode Systole allows the 96 of the cycle that is delivering to be set This value is typically about 35 which must be entered as 0 35 43 June 19 2013 Revision 1 2 In Set RATE Mode Systole is the same as in Standard Mode Systole rate is the rate in ml min that the pump is allowed to increase to reach the set delivery rate Distole rate is the rate in ml min that the pump is allowed to increase to reach the set fill rate In Set Pressure Mode constant pressure is set and the pump varies the flow to maintain the pressure setting Max rate is the maximum rate the pump is allowed Min rate is the minimum rate the pump is allowed PID Setting P Gain is the proportional gain which dictates how fast the pump is allowed to change to obtain the required setting PID Setting Gain is the integral value used to adjust the baseline and fine tune the rate at which the pump is allowed to move to reach the programmed setting In Inverse Mode the setting for the diastole and systole cycles are reversed Otherwise the parameters a
48. nation followed by the numerical value and then the units that have been assigned to that channel This display is showing a three channel DSP4000 with two channels disabled OPTICAL PROCESS MONITOR 6 2 Channel Readings Temperature and Pressure Readings Pressing the AUX key will give you a display that reads the channel designation followed by the temperature pressure values per channel These will only show real time data from an actively com pensating input source barometer or RTD probe 6 3 Channel Readings AGC Readings These readings can be useful for feedback regarding sensor effectiveness or placement in regards to the fiber optic cable High AGC readings indicate a high gain applied to the LED which can be caused by photo bleaching of the sensor chemistry or by improper alignment of the chemistry in relation to the fiber optic cable 68 June 19 2013 Revision 1 2 7 Changing expired sensors The Status LED in the upper right corner of the DSP4000 front panel provides a visual indication of the AGC or RAGC level during operation The Status LED is e Green when the AGC level is less than 3000 for all four channels e ow when the level for least one channel is between 3000 and 3500 e Red when the AGC for one channel exceeds 3500 An AGC or RAGC reading of 3500 or higher indicates a need to replace the sensing element or an obstruction or break in the optical path When the Status light is red 1 Check t
49. nts e Physiological macro to nano fluidics infusion pumps e Animal to cell electrophysiological monitoring systems e Organ to cell perfusion bath and chambers to mimic physiological conditions to study drug nutrient gene and cell therapies e Live cell imaging and perfusion chambers to better understand the affect of shear force drugs nutrition on cells and tissues e Cell engineering tools from electroporation to pneumatic injectors 5 June 19 2013 Revision 1 2 Chapter 1 Introduction The ORCA Bioreactor is the first system designed to meet the needs of the modern regenerative medicine research scientist The system is able to address the needs of both the decellularization and recellularization processes of various organs Physiological conditions can be mimicked through control algorithms that regulate flow rates profiles and pressures Critical readings can be taken from both inside the organ as well as in the support environment through the use of sensors The Concurrent Method Development software logs all modifications and operator entries along with the result that was measured with the system sensors allowing a complete review of an experiment and direct translation into a research method A method storage system makes it easy to reproduce methods and conditions across multiple experiments The image capture software allows for images to be captured in real time from cameras monitoring visible IR UV and fluorescence other sy
50. o use the 1 point calibration on the DSP4000 When complete put the DSP4000 into RUN MODE and collect data into LabChart Switch to the lower oxygen solution such as aerated buffer to capture the low value Once done stop data recording and highlight the full section of data that includes both the high and low calibration points Right click on the range in the channel and choose Units Conversion from the drop down menu Use the values displayed on the DSP4000 for the 2 point calibration in LabChart 66 June 19 2013 Revision 1 2 5 4 Carbon Dioxide Sensor Calibration A 1 point calibration should be performed on the DSP4000 when a known gas concentration e g 5 is not read accurately on the DSP Display Typically this calibration point is at 5 as that is the concentration of CO2 that is used to maintain proper pH in most bicarbonate buffers used in perfusion systems When the 1 point calibration is complete put the DSP4000 into RUN MODE and collect data into LabChart Switch to the zero gas or solution for 5 or 10 minutes respectively or until the reading has fully stabilized and continue to collect data for the low point Once done stop data recording and highlight the full section of data that includes both the high and low calibration points Right click on the range in the channel and choose Units Conversion from the drop down menu and use the values that are displayed on the DSP4000 for the 2 point calibration in LabChart
51. off Windows auto updates which could cause the computer to restart whilst running an experiment Before running the software for the first time locate the folder Harvard Data on the desktop and copy paste it into the My Documents folder as a backup This ensures that a backup copy is available in case the program is ever shut down incorrectly Click on the Harvard Data Folder on the desktop Select COPY Click on the My Documents Folder on the desktop Select PASTE Running amp Terminating the Program To run the program select the initiation icon from the Desktop It is important to shut down the ORCA controller software in the following manner Close the program using CTRL Q or the Stop option WARNING Shutting down the program by shutting off the PC or using any other method may result in a corrupted CONFIGURATION FILE and not allow the program to initiate properly the next time it is used As a result the red X button normally found in the upper right corner of the screen has been disabled 36 June 19 2013 Revision 1 2 BIO 1 File Operate Tools Notification Support The Main Screen Rate150I Run5 i Default x c 150 00 ml min 100 00 ml min Constant Rate Constant Rate Temp 2284 lt Temp 20 39 lt Press NaN mmHg Avg Press 2 60 mmHg Avg Rate 0 00 ml min Rate 0 00 ml min Temperatures Pressure FlowRate Pump iCharts Pump 2 Charts Pump 2 Pressure Avera
52. r Four Single Channel L Sipper tube Heads Combined by a Y 9hest flow rate Ventilator pathway Flow to Oxygenator Flow to Pre O sensor Flow to O5 CO pH sensor May be added to the Thermocouple pump with two dual channel heads to allow addition and removal of reservoir medium Two Dual Channel Heads Arrows indicate direction of flow 4 channel PharMed Tygon Teflon Opaque flexible Clear flexible Translucent more rigid L S 16 1 8 ID 1 8 OD 1 8 1 6mm ID w 720963 B 1721259 30358 L S 25 3 16 4 8mm 10 720964 L S 17 1 4 ID EE o5 1 4 6 4mm ID 720965 E 1721256 IP 30345 L S 18 i 3 8 ID 3 8 OD 3 8 7 9mm ID m 720967 J 721270 30346 12 7mm ID 1 2 ID 1 2 OD a 7 COOS J 721268 K 30347 Key cont next pg gt 20 June 19 2013 Revision 1 2 4 i lj Lj 1 8 clamp 30497 g connector 30357 lj 4 connector 30370 Nl 1 4 clamp 721670 Q RY 3 8 connector 721577 3 8 clamp 721671 E 9 1 2 connector 30496 1 2 clamp 30495 MM Connector g NPT Barb right angle 30511 ANIJ Connector aNPT Barb 30526 V Luer Lok Fitting 30259 Fitting Right Angle M Bubble Trap 30430 Right Angle Trap XXXXX lt Y Barbs ni AE 2 plug 30410 3
53. re the same as in Standard Mode 44 Operation Mode Please select the Start Mode vou wish Run Mode Pulsatile Cycle Set Rate Mode Systole 5 systole Rate 0 000 0 000 Diastole Rate 0 000 Ok Cancel Operation Mode Please select the Start Mode you wish Run Mode Pulsatile Cycle Set Pressure Mode PID Settings F ain systole Systole Rate 0 000 0 000 0 000 iua 0 000 Diastole Rate D Gain 0 000 0 000 OK Cancel Please select the Start Mode you wish Run Mode Pulsatile Cycle Inverse Made XT June 19 2013 Revision 1 2 In Reverse Mode the systole is operated on during the delivery phase The diastole is eee operated on during the fill phase Pulsatile Cycle Reverise Mode _ Start Mode Start Mode The user has the option to set a time delay before Please select the Start Made you wish the pump starts running at the rate at which it has StartMade been set or can opt for the start immediately setting 7 Ramp allows the user to select the speed at which Start Mode the pump will deliver There are options in increments ranging from very slow to very fast E None is also an option for type of ramping NITE RR None tra Fast Very Fast Fast Note that the maximum flow rate is a constraint set Medium Slow by the type of tubing used man 1 3 Slow Please select the Start Mode vou wish Please select the Start Mode you wish
54. reful examination but does not represent a warranty of product properties Alterations in line with technical progress are reserved This DSP4000 unit is manufactured by POLESTAR TECHNOLOGIES and supplied to you by HARVARD APPARATUS All inquiries regarding this system addition application should be directed to Harvard Apparatus or to the Harvard Apparatus Authorized Distributor from whom this unit was purchased Harvard Apparatus 84 October Hill Rd Holliston MA 01746 Phone United States 800 272 2775 Phone others 508 893 8999 eMail techsupport harvardapparatus com Harvard Apparatus has locations in Canada United Kingdom Germany Spain and France 2 Safety Note Warning the equipment is designed for use in general laboratories light industrial and office environments Operation in hazardous areas and or in a flammable atmosphere is not permitted 3 General Description Application The DSP4000 Optical Process Monitor can be configured for 1 2 3 or 4 Channels defined by the number of LED modules present in the system Units are identical in functionality 57 June 19 2013 Revision 1 2 though the total sensor capacity may differ In the descriptions below all configurations will be referred to as the DSP4000 The DSP4000 is an Optical Process Monitor designed to emit and sense light in a specific range of wavelengths for the purpose of determining the concentration of the presence of the parameter s
55. rement channel The configuration utility is also used to reassign channels to different measurements 70 June 19 2013 Revision 1 2 8 Cleaning and Maintenance Optical Sensors are not affected by ambient light when in use however they are sensitive to light and should be stored in the dark when not in use The sensors can be cleaned using MUCASOL which is preferred when using the sensors in conjunction with a Hugo Sachs Isolated Organ or Tissue Perfusion System whereby there are wetted components constructed from Plexiglas If the fiber optic cables have been detached and stored or if they are otherwise possible dirty you should use a piece of scotch tape to tap on the tip to remove particulate matter Additional in depth maintenance information can be found in Chapter 6 Care and Maintenance of the DSP4000 Optical Process Monitor User s Manual You will need the following items to inspect and maintain the DSP4000 De ionized water Chem Wipe or soft lint free cloth Scotch tape Magnifying glass Compressed air or nitrogen Adjustable wrench All Polestar sensors including sensing elements iDots peel and sticks and puncture probes are sensitive to ambient light and will photo bleach over time unless stored in a dark place between uses Bright sun light will accelerate the aging process of the sensor Photo bleaching affects the useful life of the sensing element but not the accuracy of measurements 71
56. rsal Precautions All blood products or products potentially contaminated by blood or other body animal fluids should be treated as potentially infectious materials Personal protective equipment should be worn at all times when using the In Breath Bioreactor to protect personnel from becoming contaminated as well as to help prevent cross infection and cross contamination Bench tops equipment and other potentially contaminated surfaces should be cleaned and disinfected according to the manufacturers and or the facility s procedures Any article used to clean potentially contaminated surfaces should be disposed of as Biohazardous Waste CAUTION Failure to use the manufacturers cleaning and disinfecting procedure could result in damage to the surface or equipment Biohazardous Waste Dispose of biohazardous waste according to local Regulatory requirements Accessories Manual Pressure Calibrator 73 4479 Cleaning Standard Laboratory protocols may be used In general we recommend the following steps prior to autoclaving Flushing with deionized water Washing with 70 IPA Flushing with deionized water Washing with a mild detergent Flushing with deionized water moo S Stainless steel parts may be sonicated 53 June 19 2013 Revision 1 2 Parts List Part Number Bubble Trap Ball 14 chamber 1 PolyPro bal Bubble Trap Ball 10 chamber 2 4 PolyPro ball Port Extra Chamber 14 chamber Port E
57. s 5 CO2 with balance air or 95 N2 5 CO2 TECHNICAL NOTE Oxygen Sensor calibration is always performed based on air saturation and not dissolved oxygen saturation 20 99 o DO 100 Air Saturation 95 DO 454 Air Saturation If a 2 point calibration needs to be performed then the following zero low standards should be used pH buffer pH 5 or lower this is the minimum output of the Polestar CO2 Room Air equilibrated distilled water which has a CO2 level of 0 or Nitrogen Gas O2 pO2 zero solution Catalog Number 73 3812 or Nitrogen gas saturated solution The optical sensing films consist of fluorescent indicator chemistries immobilized in either hydrophilic pH sensor or hydrophobic DO and CO2 sensors polymer membranes The gaseous O2 CO2 or ionic H species of interest move freely between the solution and interior portions of the sensing membranes to maintain an equilibrium of partial pressure O2 CO2 or chemical potential pH The time to reestablish the equilibrium condition following a change in the 63 June 19 2013 Revision 1 2 solution is dependent upon the species rate of diffusion through the membrane faster at 37 C For the purpose of calibration it is recommended that the user allow a minimum of 10 minutes exposure of cycling the calibration solution through the sensor to ensure complete equilibration of the sensing film Note it is important that the calibration solution flow over the sensor during the c
58. s are used and the fourth Is available for specialty configurations Three pumps may be initially purchased and the user may add the fourth pump on later Can the software be installed on the computer of my choosing The ORCA software is typically run from the laptop provided It can be run from a standard desktop PC as long as it is configured properly If you have a desktop we are able to configure it our laboratory If you are having an on site installation we could also configure it then Gas Monitoring System How are gas monitoring CO O2 and pH monitoring typically performed The system that Harvard currently uses is purchased from Polestar and performs with a high degree of precision and accuracy The system provides the ability to monitor CO2 and pH from a single point typically the reservoir Another ability is to measure the pre and post organ oxygen levels allowing for easy determination of the oxygen consumption rate of the organ 74 June 19 2013 Revision 1 2 Oxygenating System How are gas levels CO O controlled The typical manner to oxygenate a system is to have media from the reservoir sent through the intraluminal side of an oxygenator using one of the channels of a peristaltic pump There are a wide variety of oxygenators The D150 is used for small animal organs as it minimizes the volume and has shown the capacity needed for rodents to rabbits For large animals the higher capacity D200 is often used In the
59. s come in various sizes and with different ports to accommodate your protocols How is temperature monitored A temperature probe is normally placed in the heater reservoir to control heating functions and heat the medium when the temperature dips below the set point 15 June 19 2013 Revision 1 2 HARVARD APPARATUS REGENERATIVE TECHNOLOGY Harvard Apparatus Regenerative Technology 84 October Hill Drive Holliston MA 01746 USA www harvardapparatusregen com 508 893 8999 regenmedicine harvardapparatus com 76 June 19 2013 Revision 1 2
60. sing to view the files must have a converter pro gram installed One such program is TMS Importer which can be obtained from TMS Importer http zone ni com devzone cda epd p id 2944 or http vimeo com 22639555 x TMS Importer Plus How to Copy Attributes Alex Hoffman w TMS Importer is a tool that allows you to enter data in a spreadsheet and import it instantly into TMS It is also designed to allow you to share datasets between institubions and there s even a link to eMuseum Network that can I you work collaboratively by downloading records from a lending institution now there s no need to manually enter records for loan objects Today s webinar will cover all the interesting and useful functionality of this important new tool As a bonus we ll also show the new M s a Analog Inputs The ORCA controller allows you to feed in any analog signal and this software can capture display and save the traces This is commonly used with the PoleStar gas monitoring system Temperatures Pressure Flow Rate Pump 1Charts Pump 2 Charts Pump 3 8 Pump 4Charts Analog Inputs Analog Input 1 m oon 1 Analog Input 2 4 0195 Analog Input 3 4 vw 003 Analog Input 4 0012 0 5 Voltage 0 6 0 5 0 3 D 1 I 0 1 DN aina DAR T l lt Hig id dm
61. stems such as ultrasound ECHO are Supported 1 1 Warning and Caution Statements The use of a WARNING statement in this User Manual alerts you to a potential safety hazard Failure to observe a warning may result in a serious injury to the user The use of a CAUTION statement in this User Manual alerts you to where special care is necessary for the safe and effective use of the product Failure to observe a caution may result in minor injury to the user or damage to the product or other property 1 2 Intended Use The ORCA Bioreactor is a system used for the purposes of monitoring and studying an isolated organ it is intended for research use only Use of this device in non research settings must be conducted under local Regulatory requirements consult your local Regulatory Authority 6 June 19 2013 Revision 1 2 Chapter 2 Getting Started The following conditions must be met prior to using the In Breath Bioreactor 2 1 General Safety Requirements The ORCA Bioreactor should only be used by qualified personnel who have been trained by the manufacturer or other Authorized Representative Unauthorized use of this device is not recommended To prevent contamination aseptic procedures must be followed and personal protective equipment must be worn at all times when handling and using the Bioreactor Wherever blood products are used Universal Precautions must be followed Facility Requirements Assure that the fac
62. une 19 2013 Revision 1 2 Pumps Peristaltic Peristaltic pump heads are available in single and dual channel versions Up to four 4 heads may be used on each peristaltic pump to provide flexibility in flow range as well as the gt One Single channel Two Single Four Single channel number of channels Head channel Heads Heads required for the most 881001 881003 881010 challenging protocols High Capacity Up to 1200mL min head Mid Capacity Up to 500mL min head One Dual channel Two Dual channel One each Single and Head Heads 881004 Dual Channel Heads 881002 881005 Additional Part Numbers 881011 Mounting Screw for Two pump heads 881012 Mounting Screw for Three pump heads 881013 Mounting Screw for Four pump heads Pumps Pulsatile The pulsatile pump is used in some large animal systems to emulate ventricular action of the heart It allows for minimal hemolysis and is ideal for moving emulsions suspensions and blood 10 June 19 2013 Revision 1 2 Chambers Solid Organ Chambers Large Solid Organ Chamber Intermediate Solid Organ Small Solid Organ 14 880420 Chamber 10 880429 Chamber 5 5 880431 holds approx 32 8L holds approx 11 6L holds approx 2L Solid organ chambers are autoclavcable and are suitable for multiple species and sizes of organs Tubing sets can be changed to accommodate the wide range of flow rates The layout of the
63. urce RTD for example you can designate the RTD Probe on Channel 1 to be the Common Source and the other temperature dependent sensor on the Tri Fold Flow cell can be directed to use the Common Source If another individual sensor is located elsewhere then a second RTD probe can be installed on that channel and you would then direct the DSP4000 to use RTD as the source for that Channel If an RTD probe is not used then the temperature can be set manually 64 June 19 2013 Revision 1 2 Manual Temperature Input Manual Input allows you to specify the temperature value to use for compensation This option should only be considered for applications where there is little or no variation in temperature or where the temperature is well regulated To set the temperature manually 1 Select Manual Input in the Temperature Source menu and press ENTER to show the currently set temperature for manual input The default is 20 C The range is 20 C to 100 C 2 Press the up or down arrow to increase or decrease the setting by 0 1 C or hold the key down to rapidly change the value Press the left or right arrow keys to change the position of the cursor to the tens ones or tenths position 3 Press ENTER to accept the displayed setting and return to the Setup menu 5 1 Set Barometric Pressure Measurements taken for CO and O are sensitive to barometric pressure Unless a pressure moni tor is available with a 4 20mA analog output which
64. vailable O2 O2 Air Sat O2 PPM or PPB or SUB PPB 02 mmHg O2 deg 58 June 19 2013 Revision 1 2 pH Measurement ranges available Low Physiologic 4 7 Mid Physiologic 5 5 10 Carbon Dioxide Measurement range 0 30 Units of measurement available CO2 CO2 mmHg SMR 4 Initial Setup of DSP4000 Unit Quick start Guide Please refer to the Quick Start Guide that was supplied with your DSP4000 unit It is especially important to do the following upon initial setup Open the door to the unit loosen screws with fingers or screwdriver and do the following Remove the USB Key and plug it into your PC and copy the contents to your Desktop or preferred location on your computer Take special note of the manual recommended to print a copy if desired Return the USB Key to the Unit Verify that the toggle switch next to USB KEY is in the OFF Position before plugging in the AC Power Adapter See Figure 2 3 of DSP4000 Optical Process Monitor User s Manual Turn ON the DSP Monitor by flipping the toggle switch The Monitor should warm up for 30 minutes before first time use and at any time when powering on from a full shut down where the toggle switch inside the unit is in the off position Generally the unit can be left in Standby Mode the Main Menu is displayed between uses unless one of the following requires a full shut down e You are connecting I O as described in Section 4 Electrical Conne
65. w Command CH 1 l Flow ml m Time second This tab gives the raw and average pressure readings for Pump 2 as well as the flow rate Note UCL and LCL refer to statistical controls that the user develops when a method is validated They are able to give you a graphic representation so that the user is able to see if there are certain characteristics in the data for example a spike gradual shift or sine like wave 51 June 19 2013 Revision 1 2 4 5 Image Capture System ImageSystem vi Image Capture System seta HARVARD APPARATUS Experiment Name 3 5 2013 8 31 08 AM Experiment Name 1280x720 0 26X 32 bit RGB image 172 173 159 0 0 Note to Overlay cool Camera allows you to select which camera you want to capture on the screen There is no real limit to the number of cameras that you have hooked up Remember you can always utilize the camera on the laptop as well Lamps are used with Dino lite cameras This button turns on and off the lights on the camera displaying the left screen as live image The camera button lets you take a snap shot and save it on the right screen while still e Overlay notes allow you to add notes to the picture before you save it e Note to overly allows you to add in notes 52 June 19 2013 Revision 1 2 Chapter 5 Care amp Maintenance Cross Contamination Prevention Biohazardous Waste and Product Disposal Cross Contamination Prevention Unive
66. xtra Chamber 10 amp 5 5 chamber Probe Temperature Sensor 12 30 5 cm Probe Temperature Sensor 6 1 8 diameter Bottle Small Animal fittings 1 000 mL 890436 Bottle Small Animal fittings 2 000 mL 880437 Bubble Trap 1 2 30430 Bubble Trap 1 4 30450 Transducer Pressure Cable 880366 Tubing PharMed L S 16 1 8 3 1mm ID 120963 Tubing PharMed L S 17 1 4 6 4mm ID Tubing PharMed L S 18 5 16 7 9mm ID 120965 120967 Tubing PharMed L S 25 3 16 4 8mm ID 120964 Tubing Teflon 1 8 3 18mm OD 30358 Tubing Teflon 1 4 6 35mm OD 30345 Tubing Teflon 1 2 12 7mm OD Tubing Tygon 1 8 3 18mm ID Tubing Tygon 1 4 6 35mm ID Tubing Tygon 3 8 9 53mm ID Tubing Tygon 1 2 12 7mm ID Y Barb Fitting 3 16 4 77mm Y Barb Fitting 3 8 9 53mm Clamp 3 8 9 53mm Clamp 1 2 12 7mm Connector 1 8 3 18mm Connector 1 4 6 35mm Connector 1 2 12 7mm Quick Connector Chamber Fill amp Drain Connector 1 8 Barb 1 4 28 UNF Connector 1 4 Barb 1 8 NPT Elevator Asmbly 14 chamber June 19 2013 Revision 1 2 Appendix A PoleStar Setup and Operation Application Instructions Polestar Fluorescence based Optical Biosensors Harvard Apparatus Version 1 21 061112 Written primarily for applications involving isolated organ and tissue See manufacturer s manual for details regarding use in standard applications This guide is NOT a replacement for the manufacturer s manual 55 June 1

ORCA Bioreactor Manual - Harvard Apparatus Regenerative

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