ORCA Bioreactor™ - Harvard Apparatus Regenerative Technology
Contents
1. HARVARD APPARATUS REGENERATIVE TECHNOLOGY Harvard Apparatus Regenerative Technology 84 October Hill Drive Holliston MA 01746 USA www harvardbioscience com 508 893 8999 enmedicine harvardapparatus com Contents Symbols Overview Chapter 1 I ntroduction Chapter 2 Getting Started 2 1 Safety Requirements 2 2 Equipment Components 2 3 Heater Reservoir 2 4 Pumps 2 5 Chambers Chapter 3 Assembly I nstructions 3 1 Lung System 3 2 Heart 3 3 Aortic Valve System 3 4 Hollow Organ System 3 5 Pump Head Assembly Disassembly 3 6 Accessories Chapter 4 Operating nstructions 4 1 Sterilization 4 2 Reservoir Port Sizing 4 3 Controller Software 4 4 Decellularization 4 5 Recellularization Chapter 5 Care amp Maintenance 5 1 Parts List Appendix A Polestar Instructions Appendix B Frequently Asked Questions 12 15 16 17 18 19 19 20 21 22 29 30 31 32 33 34 52 53 55 January 22 2013 Revision 1 0 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 product 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 pr2. 0 12 1 6mm 1D eee 1 8 0D 881105 L S 17 i 1 4 ID 1 4 OD 1 4 0 25 6 4mm ID 991100 721256 30345 L S 18 1 3 8 ID 3 8 OD 3 8 G 0 31 7 9mm ID a 981101 721264 30346 j 1 2 ID 1 2 OD 1 2 E J 721268 N 30347 Clamps Connectors Y Barbs I 1 8 clamp XXXXXX P 1 8 connector xxxxxx Y 3 8 Y barb ooxxx MI 1 4 clamp 721670 Q 1 4 connector xxxxxx N 3 8 clamp 721671 R 3 8 connector 721577 0 1 2 clamp XXXXXX 9 1 2 connector xxxxxx 13 January 22 2013 Revision 1 0 Note The following is a diagram representing a configuration for the bioreactor for a arge animal lung system with cannulation of the pulmonary vein 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 idle Reservoir Port Details pg 25 t Trifold post organ 14 January 22 2013 Revision 1 0 Key Perfusion pathway into organ Oxygenation Polestar pathway Sipper tube Ventilator pathway Polestar transducer pathway f T Barb lt Y Barb ss Clamp TC Thermocouple Pumps Head Config Dual offset Dual offset 4 channel Typical Function Perfusion into organ Empty chamber to reservoir Oxygenation and analytics pathway 3 channel Prov
3. 1 0 Appendix B Frequently Asked Questions 55 January 22 2013 Revision 1 0 HARVARD APPARATUS REGENERATIVE TECHNOLOGY Harvard Apparatus Regenerative Technology 84 October Hill Drive Holliston MA 01746 USA www harvardbioscience com 508 893 8999 regenmedicine harvardapparatus com 56 January 22 2013 Revision 1 0
4. 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 events 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 perfu
5. 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 Flow Direction aie Changes the direction that the pump rotates a right arrow indicates clockwise bya Create or Select Step __Use this to create or select a Step See Step Creation on pg 24 for additional info Green Light Glows when pump is on j Step Mode r See Step Mode on pg 22 for additional info _ Start Mode See Start Mode on pg 22 for additional info a Stop Mode See Stop Mode on pg 23 for additional info z Repeat Mode See Repeat Mode on pg 23 for additional info Pump Selector Choose from peristaltic syringe pulsed blood or ventilator modes January 22 2013 Revision 1 0 Step Mode Start Mode Start Mode Please select the Start Mode you wish Run Mode Constant Rate Please select the Start Mode you wish Run Mode lt Constant Rate Constant Pressure Flow Flow Rate Direction imliy 100 E gt Flow Direction gt Pulsatile Cycle Pressure Oscillation On this screen Run Mode Flow Rate and Flow Direction can be controlled For the Run Mode parameter the pump can be set for Constant Rate Constant Pressure Pul satile Cycle or Pressure Oscillation Start Mode Start Mode Please select the Start Mode you wish StartMode Ramp lt Mone Ultra Fast Star
6. 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 the 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 e Store the cylindrical plug in a safe place Channel Terminal Labels RTD1 Black 1 RTD1 White 3 Channel 2 RTD Black 4 RTD Red 5 RTD White 6 Channel 3 RTD3_Black 7 RTO3_Red 8 RTD3_White 9 Steet tease Channel 4 RTD4_Black 10 RTO4_ Red 11 RTD4_White 12 Ae OO Me p FMA Step 5 Insert the bare wires as per the following table Clamp Pace eem down using screwdriver Connect the threaded RTD probe tip into your sensor The Flowcells and the Tri fo
7. x lt 1 to account for positive deviation Flow Rate Constraints Tubing Size Flow Rate Minimum Flow Rate Maximum ml min ov di 27 January 22 2013 Revision 1 0 Master Controls Master Controls 1 starts all pumps 2 stops all pumps 3 incrementally adjusts flow rate of all pumps 4 notepad Experiment Note Please enter your Experiment Mote below Note Number 1 Cancel Any experiment notes entered will appear on the Experiment Log Note All actions will be logged in the Experiment Log once the Master Start button is clicked Any experimental changes will continue to be logged until the Master Stop icon is clicked This is to prevent logging extraneous information while retaining valuable information about adjustments 28 January 22 2013 Revision 1 0 Heater Temperature Control amp Pressure Channel Calibration Heater Temperature Control x Heater Controls Access the Temperature Control menu through Tools gt Heater Probe To Control TZ 1 z Heater Control Probe to Control allows the user to select between four different probes labeled TC 1 to TC 4 Disabled J Heater Control can be set to either Enabled or Disabled SetPoint ee Set Point allows the user to set the temperature in degrees Celsius The Pressure Channel Calibration menu can be accessed Pressure Channel Calibration z 7 through Tools gt Calibration gt Pressure Please select the pressure cha
8. AGC level is less than 3000 for all four channels e 2 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 the 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 2 Contact Harvard Apparatus or your Harvard Apparatus Appointed Distributor for the correct re 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 1 Remove the Fiber Optic cable s 2 Using a wrench if unable to remove with finger
9. Be sure to follow your laboratory s individual protocols 21 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 January 22 2013 Revision 1 0 4 3 Harvard Apparatus Pump Software Operation I nstructions This pump 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 Pia gets Foo Honan Steet HARVARD APPARATUS THI 3 56 06 PM Yee Exparteent Time Data fat Te lial Tia Feo Daig uh 1E Eirg Te Parri Tee Lager fate een Erpereent Log tra Purrscr hay EF i l qiqiqiet 1 FS 45 fd ES Se ee OS a A A ee The Main Screen 22 January 22 2013 Revision 1 0 23 Fie Operate Tools Wotification Suppor Temp 19 95 c Press 254 86 mmHg Rate 0 00 ralmin x 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
10. and O are sensitive to barometric pressure Unless a pressure monitor is available with a 4 20mA analog output which can be fed into the DSP unit an accurate barometric 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 baro metric pressure plus gauge pressure The default value is 14 7 PSI 760 mmHg To use a manually input value for pres sure 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 Calibration Menu on DSP4000 To reach the calibration menu select Setup Mode from the Main Screen Choose the Channel being calibrated and press ENTER Th
11. bed 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 measurement channel The configuration utility is also used to reassign channels to different measurements 49 January 22 2013 Revision 1 0 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 e De ionized water e Chem Wipe or soft lint free cloth e Scotch tape e Magnifying glass e Compressed air or nitrogen e Adjustable wrench All Polestar sensors includi
12. d 881003 Mid Capacity Up to 500mL min Dual Channel mid capacity Single Head Dual Head 4 channel 881002 881004 Pumps Pulsatile Single amp Dual Channel 1 high capacity 2 mid capacity Dual Head 3 channel 881005 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 January 22 2013 Revision 1 0 Chambers Solid Organ Chambers Large Solid Organ Chamber Intermediate Solid Organ Small Solid Organ 14 880420 Chamber 10 880429 Chamber 4 5 880431 holds approx 32 8L holds approx 11 6L holds approx 1 4L Solid organ chambers are autoclavable 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 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 intralumi
13. d 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 careful 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 Operation in hazardous areas and or 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 36 January 22 2013 Revision 1 0 though the total sensor capacity may differ In the descriptions below all configurations will be referred to as the DSP4000 The DSP4000 i
14. e 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 Connections 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 38 January 22 2013 Revision 1 0 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 F PoLesTR OPTIC Al DDA ECE LATAN 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 If you wish for a full power shutoff then after the above step open the DSP4000 and flip the toggle switch to OFF Saving the Log file is optional If you have performed a calibration procedure or changed any other se
15. e 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 January 22 2013 Revision 1 0 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 facility 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 of at least BCS Level 2 Failure to provide a means to conduct a
16. en scroll down to Calibration press ENTER and follow instructions OPTICAL PROCESS MONITOR le 44 January 22 2013 Revision 1 0 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 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 uni
17. ides flexibility for the most challenging protocols Tubing Sizin PharMed Tygon Teflon g Opaque flexible Clear flexible Translucent more rigid L S 16 m 1 8 A 0 12 1 6mm 1D eee 1 8 0D 881105 L S 17 i 1 4 ID 1 4 OD 1 4 0 25 6 4mm ID 991100 721256 30345 L S 18 n A 3 8 ID 3 8 OD 3 8 G 0 31 7 9mm ID o 981101 721264 30346 j 1 2 ID 1 2 OD 1 2 E J 721268 N 30347 Clamps Connectors Y Barbs I 1 8 clamp XXxxxx P 1 8 connector xxxxxx Y 3 8 Y barb ooxxx MI 1 4 clamp 721670 Q 1 4 connector xxxxxx T Barbs N 3 8 clamp 721671 15 0 1 2 clamp XXXXXX R 5 3 8 connector 721577 1 2 connector xxxxxx T 3 8 to 1 8 T barb xxxxxx January 22 2013 Revision 1 0 3 2 Heart System Note The following is a diagram representing a configuration for the bioreactor for a arge animal heart system Your individual laboratory s setup may differ from what is depicted below 16 January 22 2013 Revision 1 0 3 3 Aortic Valve System Note The following is a diagram representing a configuration for the bioreactor for a arge animal aortic valve system Your individual laboratory s setup may differ from what is depicted below 17 January 22 2013 Revision 1 0 3 4 Hollow Organ System Please refer to the Hollow Organ Bioreactor manual for information on assembly and operation of trachea esophag
18. ior 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 distribut ed without the expressed prior written permission of the copyright holder U S international and foreign patent applications are pending 3 January 22 2013 Revision 1 0 Symbols Used emt en 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 January 22 2013 Revision 1 0 Overview HARVARD REGENERATIVE TECHNOLOGY Harvard 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
19. it is important that the calibration solution flow over the sensor during the calibration process Failure to use a flowing calibration solution can affect the calibration results 42 January 22 2013 Revision 1 0 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
20. ither 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 January 22 2013 Revision 1 0 Pumps Peristaltic Single Channel high capacity Peristaltic pump heads are available in single and dual channel versions Multiple heads may be used on each peristaltic pump to provide flexibility in flow range as well as the number of channels required for the most Single Head challenging protocols 881001 High Capacity Up to 1500mL min Dual Offset Hea
21. ividual sterilization protocols Pharmed tubing may be used throughout the system As a cost saving measure or to have tubing that is transparent Tygon tubing may be used see illustrations If the flow path is autoclaved 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 tubing that was on the barb is cut and discarded and that the fresh tubing is slid onto the barb Ty wraps may also be used on barb fittings 20 January 22 2013 Revision 1 0 4 2 Reservoir Bottle Configuration amp Typical Usages 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 laboratory 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 control 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
22. ld Flowcells both have built in threaded ports to accommodate the RTD probe 40 January 22 2013 Revision 1 0 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 SENSOR HOUSING PS v A N DISPOSABLE pH COs O2 SENSING ELEMENT 4 4 Connecting DSP4000 to Data Acquisition System e f 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 Data 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 Plea
23. lly contaminated surfaces should be disposed of as Biohaz ardous 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 Peristaltic pumps with isolated heads capable of being used in a CO incubator Cleaning Flush all fluid paths with deionized water and either follow with or wipe with 70 IPA Stainless steel parts may be sonicated 32 January 22 2013 Revision 1 0 Parts List 33 Amount Equipment Clamp 1 8 3 18mm Clamp 1 4 6 35mm Clamp 3 8 9 53mm Connector 1 8 3 18mm Connector 3 8 9 53mm PharMed tubing L S 16 0 12 3 1mm ID PharMed tubing L S 17 0 25 6 4mm ID PharMed tubing L S 18 0 31 7 9mm ID PharMed tubing L S 25 0 19 4 8mm ID Teflon tubing 1 8 3 18mm OD Teflon tubing 1 4 6 35mm OD Teflon tubing 3 8 9 53mm OD Teflon tubing 1 2 12 7mm OD Tygon tubing 1 8 3 18mm ID Tygon tubing 1 4 6 35mm ID Tygon tubing 3 8 9 53mm ID Tygon tubing 1 2 12 7mm ID Y Barb 3 8 9 53mm 3 8 to 1 8 T fitting ENTAO XXXXXX 881105 881100 721264 January 22 2013 Revision 1 0 Appendix A PoleStar Setup and Operation Application Instructions Polestar Fluorescence based Optical Biosens
24. located in different areas then multiple RTD probes can be used A Channel is instructed to look at either a Common Source 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 43 January 22 2013 Revision 1 0 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
25. n the driveshaft This will prevent any buildup of evaporated salt crystals from forming on the 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 For the hollow organ bioreactor should I sterilize using plasma sterilization or would autoclaving 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 pre vent leaks It should not be necessary to remove these fittings from the Teflon because of the steri lization 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 53 January 22 2013 Revision 1 0 Appendix B Frequently Asked Questions 54 January 22 2013 Revision
26. nal 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 January 22 2013 Revision 1 0 Chapter 3 Assembly I nstructions 3 1 Lung System Note The following is a diagram representing a configuration for the bioreactor for a arge animal lung system without cannulation of the pulmonary vein 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 Reservoir Port Details pg 25 12 January 22 2013 Revision 1 0 Key Perfusion pathway into organ Oxygenation Polestar pathway Sipper tube Ventilator pathway lt Clamp Y Barb TC Thermocouple Pumps Head Config Dual offset Dual offset 4 channel Typical Function Perfusion into organ Empty chamber to reservoir Oxygenation and analytics pathway 3 channel Provides flexibility for the most challenging protocols Tubing Sizin PharMed Tygon Teflon g Opaque flexible Clear flexible Translucent more rigid L S 16 m 1 8 A
27. ned in section 3 1 of this document 3 1 Measurement Specifications Oxygen Measurement ranges available High 0 500 Air Sat Low 0 20 Air Sat Units of Measurement available 02 O2 Air Sat O2 PPM or PPB or SUB PPB O2 mmHg O2 deg pH 37 January 22 2013 Revision 1 0 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 pre ferred 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 b
28. ng 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 50 January 22 2013 Revision 1 0 Troubleshooting Guide The DSP4000 does Insufficient or too much Disconnect power from the not display power is being applied to DSP4000 measurements or the DSP4000 Verify that the voltage is 24 VDC els 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 gt 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 Check the AGC for each mode or kicks out the maximum value ete a a channel shortly after because RENAD There is a poor optical anil a ara The Status LED is connection red A 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 Peus 2 Clean the optical surfaces using e 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 a
29. nnel 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 O02 CO2 pH Parameters These are displayed as in the image to the left The channel designation 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 compensating 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 47 January 22 2013 Revision 1 0 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
30. nnel you wish to calibrate e First select the desired Pressure Channel given Eese cher sf The Filter may be set to either Mean or Raw The Actual Value is the instantaneous value for the Actual Value pressure reading Cal value Average Value Average Value is the average reading over a few Calibration z F i j D D a seconds The 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 along with the Calibration button to calibrate pressure readings 29 January 22 2013 Revision 1 0 Decellularization 30 January 22 2013 Revision 1 0 Recellularization 31 January 22 2013 Revision 1 0 Chapter 5 Care amp Maintenance Cross Infection Prevention Biohazardous Waste and Product Disposal Cross Infection Prevention Universal Precautions All blood products or products potentially contaminated by blood or other body animal fluids should be treated as potentially infec tious 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 manufac turers and or the facility s procedures Any article used to clean potentia
31. nue 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 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 46 January 22 2013 Revision 1 0 6 Viewing Data on DSP4000 Run Mode will by default show you the channel measurement data for the sensor plugged into that cha
32. on In these situations it is common to use some 5 CO2 gas mix for the 1 point calibration of the CO2 sensor such as 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 9 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 oH 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 02 CO2 or chemical potential pH The time to reestablish the equilibrium condition following a change in the 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
33. ors 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 34 January 22 2013 Revision 1 0 Table Of Contents 1 Introduction Manufacturer s details 2 Safety Note 3 General Description Application 3 1 Measurement Specifications 4 Initial Setup of DSP4000 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 35 January 22 2013 Revision 1 0 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 shoul
34. s 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 correct re placement part number for your sensor type i e O PPB O PPM O Sub PPB CO pH 5 5 10 or 4 7 48 January 22 2013 Revision 1 0 SENSOR HOUSING ee E A lt P i p 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 descri
35. s 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 perfusate media 51 January 22 2013 Revision 1 0 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 REPLACEMENT QUARTZ 0 PPB SENSOR INSERT FOR SINGLE QUARTZ FLOWCELL MOUNT 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 52 January 22 2013 Revision 1 0 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 o
36. s 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 for which the connected sensor s is are configured pH O2 CO2 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 O2 CO2 A dual channel system can be configured for measurement of 2 of the same parame ters requiring only one purchase of the firmware for that measurement or 2 different parameters as long as the firmware for the parameters of interest are both purchased or 3 different 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 applications or who will be calibrating their sensors in the ranges outli
37. se connect to the channel on which you wish the particular reading to be recorded e The output range of the analog output is as follows kaiti eae amaz Minimum measurement 4mA is 2V output gt Scale read for the channel from mmHg Ommhg S000 mmHg DSPCALIB DAT in the USB KEY Factor 2 Og Oy om and Factor 3 for channels 2 and 4 in 0 100 i5 Alr Sat 0 Saturation 100 Saturation i z PPM 0 PPM 5000 PPM High scale value Dey 160 180 read for the channel PPB Dissolved Oo PPB 5000 PPS l e a I 0 Pen face channel 3 in Deg 180 180 Figure 4 9 PPB Dissolved o PPB 5000 PPB SMR 3 oe 0 PPa eo am iso e Maximum measurement 20m lt A is 9 9V 41 January 22 2013 Revision 1 0 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 pH 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 If 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 calibrati
38. septic cell processing may result in harmful contamination 7 January 22 2013 Revision 1 0 2 2 Equipment Components Store the ORCA Bioreactor in a cool dry place free from dust and other potential contaminants until ready to use Assure that the ORCA Bioreactor package was received completely and without damage if the package arrived as damaged contact your local technical support group Ensure that all parts the ORCA Bioreactor package and its peripheral devices are in good condition Do not use damaged parts January 22 2013 Revision 1 0 Heater Reservoir The heater is used to maintain temperature throughout the system to emulate physiological conditions The reservoir holds a volume of 0 5L 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 12 temperature monitoring probe There are two different sizes of bottles large and small flow that have different size ports to accommodate a wide variety of tubing and flows 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 All 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 e
39. sion 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 January 22 2013 Revision 1 0 Chapter 1 I ntroduction 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 systems 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 or patient The us
40. t Immediately Very Fast Fast Wedium Slow Very Slow Ultra Slow This screen allows the user to choose the Ramp during start up of the pump giving a range of options from ultra slow to ultra fast Selecting no ramp is an option The user can also toggle the Start Mode between starting immediately and starting with a delay 24 January 22 2013 Revision 1 0 25 Stop Mode Stop Mode Please select the Start Mode you wish Stop Mode lt Stop Never User Stop Stop on Time Target This screen gives the user the option to have the run stop automatically after a specified amount of time has elapsed The default setting is Stop Never which continues the run until the user manually terminates it Repeat Mode Start Mode Please select the Start Mode you wish RepeatMode lt Mo Repeat Repeat Till User Stop Repeat Number of Times Repeat Increase n Times On this screen the user can choose between No Repeat Repeat Till User Stop Re peat Number of Times and Repeat Increase n Times January 22 2013 Revision 1 0 Step Creation Create or Select Step Would you like to Create a New Step with your present configuation Create Step Name Rate loorUsrstp Select Select StepDescription Start Immediately Run a Constant Rate 100ml thin Infuse Direction Until User Stop Cancel Create or Select Step x would you like to Create a New Skep with your present configuation Crea
41. te Select Select Select Sten J Manual Testi Test2 Tests Test4 Tests Testo Test PressoOIUsrstp Test 1 T teste j Ratel25 UsrStp Dly2RabelOoow RunSkpkso Default best 1 iv This window allows for the creation of a New Step or the selection of a previously used one If the create box is checked a New Step can be created with input boxes for the Step Name and a brief description If the select box is checked the user may select from the drop down menu any of the previously created presets 26 January 22 2013 Revision 1 0 Pump Calibration Fie Operate Tools Wotificatian Suppor To access the pump calibration menu click on the gt image of the pump that you wish to calibrate From H this menu the correct tubing size can be selected and the calibration factor can be adjusted Pump Calibration Pump 1 0 00 rolfmin Please select tubing size and or add on calibration Factor 19 95 cog Tubing Size Calibration Factor 54 86 mmHg Lis 17 A Flow Rates Min 0 84 miroir Max 40 mlfrnin The calibration factor is selected based on the deviation from optimal flow rate In practice the user may run at a specified rate into a graduated cylinder in order to test the actual volume delivered If the volume delivered is less than the expected amount the calibration factor can be set at a value greater than 1 to compensate for this deviation Likewise it can be set at a value 0 lt
42. ts 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 to 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 45 January 22 2013 Revision 1 0 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 conti
43. tting or recorded measurement data onto the log file it is advisable to save the log file 4 1 USB Key It is important to identify which sensor was assigned to each channel in order to 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 This is done when you wish to review the log file or update lot calibration information such as 39 January 22 2013 Revision 1 0 b 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
44. us systems 18 January 22 2013 Revision 1 0 Pump Head Assembly Disassembly 3 6 Accessories Dino Lite Cameras Refer to the Dino Lite manual for instructions For regenerative medicine applications please see Appendix A for information regarding setup and calibration For full manufacturer instructions please refer to the PoleStar manual at http polestartech com support pdf dsp4000 user manual pdf 19 January 22 2013 Revision 1 0 Chapter 4 Operating I nstructions 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 procedure in its entirety prior to using the de vice Personnel should train on all processes and equipment prior to operating the ORCA Bioreactor Failure to follow aseptic techniques and failure to train on all processes and procedures prior to using the In Breath 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 One possible 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 ind
Download Pdf Manuals
Related Search