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ORCA ™ Bioreactor Manual

1. Rise Time Steady State Error 1 0 5 0 0 i i 1 i i 1 i i 1 i i i N 00 02 04 06 g i Le 14 15 i8 20 22 24 26 AB 30 Time seconds The following table may be of use when tuning the PID controller Effect of increasing a parameter independently Error Decrease Increase Small change Decrease Degrade Decrease Increase Increase Eliminate Degrade Ka Minor change Decrease Decrease No effect Improve in theory if Ka small 80 Nov 01 2013 Revision 1 4 1 Bioreactor PID Temperature Controller A HART PID Controller with blanket heater can be ordered as an accessory in order to regulate temperature in either the reservoir and or pictured below in the chamber 81 Nov 01 2013 Revision 1 4 1 Setup Instructions Select the appropriate power cord and insert into the power cord plug NOTE THE REVISION 1 PID CONTROLLERS ARE 100V ONLY The unit should ONLY be plugged into a 220V 100V transformer if the line voltage is 220V Do not connect temperature probe until you have finished setting thermocouple type and Set Point temperature Select the appropriate ORCA temperature probe and plug the connector into the port using the color coding All four types of temperature probes will work with this unit 12 30 5cm L lg 32cm OD 12 30 5cm L I16 16cm OD 6 15 25cm L Ie 32cm OD 6 15 25cm L t i 32cm OD X X X x 82 Nov 01 2013 Revision 1 4 1 Progra
2. 1 de Perfusion pressure pass through connections Perfusion inlet ports Utility ports 34 Nov 01 2013 Revision 1 4 1 Cannula Adapter Kit Cannula Adapter Kit for 5 5 Chamber 31116 for 5 5 Chamber 31117 typically for rodent lung typically for rodent heart Cannulae are available in various sizes Typical sizes include Mice 1 mm Rat 2 3 mm Rabbit 3 mm or 4 mm 35 Nov 01 2013 Revision 1 4 1 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 deform around the barb during an autoclave cycle and can become loose and allow 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 individua
3. HARVARD APPARATUS REGENERATIVE TECHNOLOGY Harvard Apparatus Regenerative Technology 84 October Hill Drive Holliston MA 01746 USA www HARTregen com 774 233 7300 cservice HARTregen com Contents Contents 2 Disclaimer J J A J 3 LAT Symbols 1 1 1 4 Overview 5 Chapter 2 Getting Started 2 1 Warning amp Caution Statements 7 2 2 Intended Use 7 2 3 Safety amp Facility Requirements 7 2 4 General Equipment Components 3 1 Identifying Components i ibrati 55 56 Chapter 5 Care amp Maintenance 57 5 1 Cleaning 57 58 i 61 icati 79 2 3 3 N 14 22 24 25 25 6 1 6 7 A 5 2 Ordering Information Replacement Part Numbers Appendix A Polestar Operations Appendix B PID Controller Theory amp Application Appendix C FAQs amp Troubleshooting UPDATE LOG Nov 01 2013 Revision 1 4 1 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 HART 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
4. 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 504000 does The AGC for at least one Verify all optical connections not go into Run channel has reached 4000 Check the AGC for each mode or kicks out the maximum value shortly after because startup channel Replace sensing elements as e There is a poor optical needed 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 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 perfusate media Ordering Information 31165 O Replacement Kit sensor glass window amp O Ring 31166 pH Replacement Kit sensor glass window amp O Ring 31167 CO Replacement Kit sensor glass window amp O Ring O Replaeementsensor 3116
5. very fast response time with respect to error Run the system and track data for about 5 minutes If the system seems too unresponsive the correct course of action would be to slowly increase the P value If there seems to be too much overshoot the I value should be lowered There is no easy answer to finding the optimal values for these parameters it is simply a matter of adjusting them based on observations and testing When in doubt typically conservative i e lower values for I and P are preferred 79 Nov 01 2013 Revision 1 4 1 Commonly used terms in PID systems Set Point the desired value for a variable at a point in time Process Variable the actual measured value of that variable Error the difference between the set point and the process variable at a given time Tuning optimizing each of the three parameters such that rise time overshoot settling time and steady state error are minimized Steady State the final value that the system achieves in which an equilibrium is maintained Steady State Error the final difference between the process variable and set point ideally zero Rise Time the time that is takes the system to go from 10 to 90 of the steady state value Overshoot the amount that the process variable goes above the final value Settling Time the time required for the process variable to settle within 5 of the final value 1 5 _Proosss Variable LI Percent Overshoot Set Point E in Time
6. PID controller feedback 46 Nov 01 2013 Revision 1 4 1 In Pressure Oscillation Mode the system alternates between using the high rate to obtain the high pressure target and using the low rate to obtain the low pressure target The pressure targets are in units of mmHg and the rates are in ml min In Pulsatile Cycle Mode the program allows for a pulsatile flow to be provided by the peristaltic pumps There are five modes of operation Standard Set Rate mode Set pressure mode Inverse mode Reverse mode In Standard Mode the parameter BPM allows the beats per minute to be set Systole allows the of the cycle that is delivering to be set This value is typically about 35 which must be entered as 0 35 47 Please select the Start Mode you wish Run Mode Pressure Oscillation StandardMode High Press Target mmHg 100 High Rate ml min 0 000 Operation Mode Please select the Start Made wou wish Run Mode Pulsatile Cycle Systole 95 J Standard Mode Set Rate Mode Set Pressure Mode Inverse Mode Reverise Mode Please select the Start Mode you wish Run Mode Pulsatile Cycle Standard Mode Nov 01 2013 Revision 1 4 1 Operation Mode In Set RATE Mode Systole is the same as in Sta nda rd Mode Please select the Start Made wou wish Run Mode Pulsatile Cycle Set Rate Mode Pi Systole rate is the rate in ml min that the pump is allowed to increase to rea
7. bench space It is recommended that tubing be fed into the pumps prior to stacking 14 Nov 01 2013 Revision 1 4 1 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 31108 Single Channel 31107 Double Channel Pump head mounting screws mounting hardware for either 2 3 or 4 pump heads 6 Connect heater power cord to the controller into heater slot 1 Add liner to heater and wrap it over the top of the heater Then place reservoir bottle inside heater Only the 4L bottle will fit snugly An extension cable 31111 is available for order if more length is needed to connect the reservoir heater to the controller 7 The smaller pronged end of the temperature probe adapters should be plugged into the appropriate port on the ORCA controller find the labeled blue slots For the reservoir heater this is typically port 1 Connect the leads on the end of the thermocouple to the corresponding female slots on the adapter cable Note Up to 3 probes can be used with an ORCA 3 1 controller system with a pulsatile blood pump The color on the probe connector should be matched with the same color on the ORCA controller panel copper to copper and silver to silver 30961 Mini T Adaptor 31079 12 Probe 1 8 OD 31078 6 Probe 1 8 OD 0 An extension cord may be needed Check the probe he
8. channels that you would like to have plotted 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 1 Charts Pump 2 Charts Pump 3 Charts Pump 4 Charts Analog Inputs E in 1 Temperature 1 ra e Temperature 2 Iv Temperature 3 n m Temperature 4 ant W Temperature c l l l l See 77 ENIM CHE ale Al NS E SRF A 125 l2 11 5 I Wmi E dhs cil 135 11 145 15 Time seconds Temperatures Pressure Flow Rate Pump 1Charts Pump 2 Charts Pump 3Charts Pump 4 Charts Analog Inputs Pressure 1 a Iv Pressure 2 n r Fressure 3 n Pressure 4 rd Pressure mmjha l l l Ern F IS ro 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 52 Nov 01 2013 Revision 1 4 1 Note If 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 box es next to Pressure 1 Pressure 2 etc Temperatures Pressure Flow Rate Pump 1 Charts Pump 2 Charts Pump 3 Charts Pump 4Charts Analog Inputs fl Pump 1 Pan Iv
9. contents or product This product is for RESEARCH USE ONLY Copyright O 2013 Harvard Apparatus All rights reserved ORCA Bioreactor is a trademark of HART HART 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 Nov 01 2013 Revision 1 4 1 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 Nov 01 2013 Revision 1 4 1 Overview HARVARD APPARATUS REGENERATIVE TECHNOLOGY Harvard Apparatus Regenerative Technology HART 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 we worked closely with leading global researchers to produce products with the highest levels of performance quality and support necessa
10. depicted below this diagram is not a replacement for your laboratory s protocols KEY 14 PharMed tubing 1 16 1 6mm ID 3 16 4 8 OD ae 16 PharMed tubing 1 8 3 1mm ID 1 4 6 4mm OD 25 PharMed tubing 3 16 4 8mm ID 5 16 7 9mm OD m oS T T fitting Z ON Single Channel Pump Head Zo ON Dual Channel Pump Head Peristaltic Pump 2 LS Organ Chamber SN ZZAN Inlet Outlet Adaptor for 500mL 1L amp 2L Reservoir Bottle Allows more tubing connections to be made Clear Nut 31141 T Fitting 31138 Green Nut 31142 31143 Note Typically green caps indicate inlets to the reservoir red caps indicate outlets 31 Nov 01 2013 Revision 1 4 1 5 5 Chamber Lid Mini Bubble Trap for 5 5 Chamber with Luer Fittings 31200 Nov 01 2013 Revision 1 4 1 Fill from reservoir Right Atrium ventricular Reservoir Pressure pressure Extra probe ports Temperature probe Left Atrium ventricGlar pressure Right Atrium ventricular Left Atrium v ntricular gt E Bubble trap empty port Rod mounting for manipulator holder 33 Nov 01 2013 Revision 1 4 1 Lung gt T A 0 d ta lid a il tel Perfusion pressure pass through connections Perfusion inlet ports Utility ports Heart HEER 7 MES
11. ml min Constant Rate Constant Rate Constant Rate Temp 2284 c Temp 290 39 sc Press NaN C NaN mmHg Avg Press Rate 0 00 ml min 2 60 mmHg Avg Rate 0 00 ml min Rate 0 00 ml min Temperatures Pressure Flow Rate Pump 1Charts Pump 2 Charts Pump 2 Pressure Average M 4 2 49 mmHg Ava Press v gt 40 00 ml min Pulsatile Cycle 606 Temp NaN ec 0 05 mmHg Avg Rate 0 00 ml min Pump 3 Charts Pump 4Charts Analog Inputs 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 Wy Experiment Log AN M 3 75 UCL gs v 3 25 Q 3 Pump 2 Pressure Raw 2 75 ta 2 5 dz 2 25 400 A Flow Command CH 1 w 5 z 200 300 Lb 60 58 56 54 2 0 B B 44 2 MD B 36 24 2 30 B B 24 2 20 18 16 14 312 310 8 6 51 Nov 01 2013 Revision 1 4 1 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
12. the case of large animal porcine or human hearts an average of approximately 5L min must be achieved generally This means that typically 10 11L min is needed during systole requiring a large pulsatile pump This pump operates by filling a chamber and a piston drives it in one pulse to the system The flow rates are constrained by the type of tubing that is used The smallest tubing 1 32 ID allows flow rates from 0 018mL min and the 3 8 ID tubing allows up to 4 8L min per pump The pulsatile pump is often equipped with 1 2 ID tubing 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 pumps 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 For applications involving large animal organ perfusion a pulsatile pump is typically used in order to achieve the necessary volume 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 the user wishes to use a desktop HART may configure it in our laboratory If an on site installation is planned it may be able to be configured then 86 Nov 01 2013 Revision 1 4 1 Heater Temperature Regula
13. the desired temperature in SetPoint degrees Celsius 38 The Pressure Channel Calibration menu can be accessed through 7oo s Calibration Pressure Pressure Channel Calibration Pressure Channel Calibration Select the desired pressure channel You can have up to four channels Please select the Input channel you wish to calibrate Calculated Measured Value 186 3908 Calculated Measured Value is the instantaneous value for the Um pressure reading zero crate 2 n ul 7 Use the Zero button to zero the actual value reading and Cal iv il MW 1 N 1600 Value along with the Calibration button to calibrate pressure s readings The zero is normally set by opening the transducer to air 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 able to use it Manual Pressure Calibrator 31075 55 Nov 01 2013 Revision 1 4 1 4 5 Image Capture System ImageSystem vi E UE Image Capture System seta HARVARD APPARATUS Experiment Name 3 5 2013 8 31 08 AM Experiment Name Captured Image Recalled Image 1280x720 0 26X 32 bit RGB image 172 173 159 0 0 Note to Overlay cool Camera 0 Camera allows you to select which camera you want to capture on the screen Enable 1 There is no real limit to the numb
14. 0 AM Experiment Name Experiment Name 1 1 Status Running Experiment Log Experiment Time Data Start Time 3 5 2013 10 38 12 AM All data will be continuously entered into the experiment log until the user stops the run Total Elapsed Time 00 00 08 Total Run Time 00 00 08 Running Time 00 00 08 Paused Time 00 00 00 Experiment Notes Logging Rate 1 second lt i Clicking on the Notepad icon allows the Experiment Log user to enter an experiment note Time Function Pump Value 4 3 5 2013 10 38 09 AM PumpOFF 1 14400 3 5 2013 10 38 09 AM PumpOFF 2 100 00 35 201310 38 2AM Start 10 38 12 3 5 2013 PumpON 1 14400 3 5 2013 10 38 12 PumpON 2 100 00 Ld Please enter your Experiment Note below Mote Number 1 42 Nov 01 2013 Revision 1 4 1 43 Rate 1501 Runs uus Constant Rate Temp 25 39 scc Press 2 60 mmHg Ava Rate 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 An 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 Flow Direction aa Changes the direction that the pump rotates a right arrow indicates clockwise Create Step Use thi
15. 31142 31076 Nut Luer Red 25 each 31143 31077 Nut Thumb for Chambers 5 each 31145 31179 30875 0680009101 Pipe Barb 1 4 1 4 NPT 25 each 31133 068000920 3108 0680009301 30410 0680009501 30470 0680009601 5 m 31075 Cable ORCA Power Italy 0680009701 Cable ORCA Power Switzerland 0680009801 Ze EDES 31080 31072 Probe Temp Sensor 12 30 5 cm x g OD 31079 31086 Probe Temp Sensor 12 30 5 cm x 4e OD 30960 31116 Probe Temp Sensor 6 15 24 cm x g OD 31078 21117 Probe Temp Sensor 6 15 24 cm x s OD 30959 30240 3096 30260 31061 30320 31062 31085 31063 31084 31064 31083 31065 Clamp 1 8 3 18mm pk 25 31087 21066 31088 31099 31089 31105 Clamp 1 2 12 7mm pk 25 31091 31104 Connector 1 8 3 18mm pk 10 31092 31100 NDE 6 35mm pk 10 31093 31067 Connector ch 9 53mm pk 10 31094 31068 Connector 1 2 12 7mm pk 10 31095 Quick Connector Chamber Fill amp Drain 3 8 31211 Quick Connector Chamber Fill amp Drain 1 4 31210 30532 30407 58 Nov 01 2013 Revision 1 4 1 Parts List cont Spring for 14 Chamber Heater 31209 Spring for 10 Chamber Heater 2ea 31207 Spring for 5 5 Chamber Heater 2 long amp short 31208 Transducer Pressure Pendotek 10ea 31074 31073 30685 30690 Tubing PharMed 1 32 0 8 mm ID 25ft 31101 31102 30340 30339 30338 30337 31103 30552 Tubing PharMed 3 4 19 1mm ID 5ft 30553 Tubing Teflon 1 8 3 18mm OD 1ft 30358 30345 30
16. 346 30347 30341 30342 30343 Tubing Tygon 1 2 12 7mm ID 30344 Y Barb Fitting 3 16 4 7 7mm 10ea 31124 Y Barb Fitting 1 4 6 4mm 10ea 31125 Y Barb Fitting 3 8 9 53mm bea 31126 59 Nov 01 2013 Revision 1 4 1 Pulsatile Pump Head 31150 31156 Pulsatile Pump grease 31164 Part No 31151 31152 31153 31154 31155 31156 31157 31158 31159 31160 31161 31162 31163 Replacement Part Cylinder Head Valve Nut Coupling Nozzle Suction Body Valve Nozzle Discharge Piston Shaft Connecting O Ring Parker 2 140 O Ring Parker 42 133 O Ring Parker 2 24 Set Screw 8 32 x 3 16 Sphere 3 4 D N 31154 60 Nov 01 2013 Revision 1 4 1 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 61 Nov 01 2013 Revision 1 4 1 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
17. 8 O Replacement sensor 31175 BUFFER SOLUTION KIT pH 6 7 41 8 500ml EA 31176 Trifold Connections Kit 1 16 with sensors 78 Nov 01 2013 Revision 1 4 1 Appendix B PID Controller Theory amp Application This section is intended to provide additional information about the theory and application of a PID control loop PID control uses a feedback mechanism to minimize disturbances and keep the system close to the user defined set point at any given time The set point can be a constant a linear function or even a sinusoidal function PID control is named after its three correcting terms proportional integral and derivative The weighted sum of these three parameters calculate the output of the PID controller Put simply P depends on present error I on the collection of past errors and D is a prediction of future error The ORCA uses PID control to regulate pressure After measuring the pressure and calculating the error error set point actual value the controller decides when to change the flow rate and by how much For example if the current pressure reading is 60mmHg and the desired set point is 3B0mmHg the controller will increase the flow rate in order to make up the difference If the set point is instead 120mmHg the controller will increase the flow rate more rapidly This is known as proportional control P If the desired pressure is not being reached quickly enough the controller may try to speed up the pr
18. Connector a NPT 2 8 Barb right angle Connector sNPT 2 8 Barb v Luer Lok Fitting 10 ea 31136 Bubble Trap 2 14 chamber Right Angle Fitting 14 chamber 1 5 Universal Plug 1 4 Barb 1 2 NPT 25 ea 31128 1 3 Barb 4 28 UNF 25 ea 31129 1 4 Y barb 31124 Y barb 0e 31125 Je Y barb Gee 31126 Bubble Trap 4 10 chamber 1 4 barb g Right angle 25 ea 31132 1 4 barb 1 4 NPT 10 ea 31133 10 ea 31127 10 ea 31131 30297 30420 30410 30450 l s pipe 18 NPT Adapter 10 ea 31134 Female Luer 1 4 28 10 ea 31136 Swabble Port 25 ea 31081 Pulsatile Pump head Reducing Connector 31150 J barb Barb 10 ea 31137 Nov 01 2013 Revision 1 4 1 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 Front View Tubing size Lateral View Tubing na a 00 ma CoD pug ne Gem EEUU 25 owammo L 07 O nm BS me Q e O mmm o Used for connections not for use in the peristaltic pump Note When feeding the tubes through a peristaltic pump it is recommended that you set a convention for flow direction that you follow for all pumps
19. 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 pum dn 71 Nov 01 2013 Revision 1 4 1 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 Inpu
20. 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 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 65 Nov 01 2013 Revision 1 4 1 Refer to Quick Start Guide for connection of Fiber Optic Cables NOTE You must
21. Probe 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 5 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 cor rect replacement part number for your sensor type amp O2 PPB O PPM O Sub PPB CO pH 5 5 10 or 4 7 Nov 01 2013 Revision 1 4 1 SENSOR HOUSING x P N m 1 DISPOSABLE pH CO2 2 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
22. Pump 2 Pump 3 Pan Iv Pump4 4 v 5 5 e 6 4 5 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 i Charts Pump 2 Charts Pump 3Charts Pump 4Charts Analog Inputs Pump 2 Pressure Average a 4 Pump 2 Pressure Raw ir M 3 75 Variance 100 UCL Pressure mm h P La tn in in l LOL de 8 LN Flow Command CH 1 n Flow ml min 8 8 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 53 Nov 01 2013 Revision 1 4 1 Reading Data Files In order to read data logged as an Excel file the computer that you are using to view the files must have a converter program installed One such program is TMS Importer which can be obtained from http zone ni com devzone cda epd p id 2944 or http vimeo com 22639555 9 GallerySystems Collect Manage Share TMS Importer 2 TMS Importer Plus How to Copy Attributes Alex Hoffman Ihe new TMS Importer is a tool that allows you to enter data in a spreads
23. This can be done with a premixed tank of gasses or a gas mixer The OKO system allows the user 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 87 Nov 01 2013 Revision 1 4 1 Pressure Transducers How many pressure transducers can be used with the system What are their capabilities The chamber is built to allow for up to four pressure transducers per ORCA system There are multiple types of pressure transducers that can be used with the system Typically single use Pendotek pressure transducers are utilized These are not autoclavable although some users have reused them following EtO or chemical sterilization It is currently unclear how long the units last after being cleaned and sterilized The peristaltic pumps can be used in a constant pressure mode or pressurized for negative ventilation Troubleshooting ORCA Software Problem An error message is displayed when using the ORCA software e From the Notifications menu click Status then uncheck the Enable box Next click Save and use the check mark to exit the screen Problem The ORCA software immediately closes upon start up e Check the following e Windows auto updates is turned off otherwise it could cause the computer to restart whilst running an experiment e The ORCA power cord is plugged into the ORCA controller and the wall e The Com
24. actor is selected based on the deviation from optimal flow rate The user may wish to run their perfusate 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 one to compensate for this deviation MEN Used for connections not for use with peristaltic pumps Specifications for PharMed BPT Pump Tubing Tubing Size 13 14 16 25 17 18 82 ml min for single pump head 0 018 18 0 63 63 2 4 0 5 1 510 8 4 840 11 4 1 140 20 0 13000 Inner Diameter in mm 0 03 0 8 0 06 1 6 0 12 3 1 0 19 4 8 0 25 6 4 0 31 7 9 0 5 12 7 Barb Size in mm Mag 1 6 lAg 1 6 1 8 3 2 She 4 8 1 4 6 4 3 3 9 5 1 3 12 7 Outer Diameter in mm 5 32 3 9 3 4 19 1 Max Pressure continuous psig bar 10 0 7 Max Pressure periodic psig bar 15 1 0 Use in Single Channel Head YES YES YES YES Use in Dual Channel Head YES YES YES YES NO 45 Nov 01 2013 Revision 1 4 1 Operation Mode Operation Mode On this screen Run Mode Flow Rate and Flow Direction can DM I UNE lied she EN een Pressure Oscillation Flow Pulsatile Cycle Direction 570 Er Run Mode There are four options to choose from e Constant Rate provides a constant perfusion rate e Constant Pressure pump alters its flow t
25. ad connector to determine if you need a large oval extension cable or the small slot extension cable 30961 Probes with Mini T connectors 30960 12 Probe 1 16 OD Mini T 30959 6 Probe 1 16 OD Mini T 31080 Intra organ probe Mini T 15 Nov 01 2013 Revision 1 4 1 8 Loosen the compression fitting then add temperature probe to reservoir Be sure not to have the probe touch the bottom of the reservoir Insert additional probe into chamber for monitoring temperature within the chamber For internal organ temperature measurements a flexible implantable probe 31080 can be utilized Note Refer to the FAQ section of the manual for more information regarding temperature probes 9 To set up pressure readings mark the cables on the connector with the number of the channel that they represent if not already labeled Connect the other end of the pressure transducer cables to the ORCA controller 10 When the system is to be used the 4 to 1 transducer cable will be connected to each of the pressure transducers see Operations section for calibration procedure It is necessary to apply a lot of pressure when screwing the Pendotek pressure transducers into the side ports on the chamber lid There are 3 side ports on the 5 5 chamber there are 4 side ports on the 10 amp 14 chambers A swabable port should be attached to each of the transducers to maintain sterility see bottom left image Straight Fitting
26. 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 OAM A POLESTAR S Technologies Inc STATUS CYPTIC Al DDANCECC MANUTAN 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 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 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 c
27. ally users may wish to add a second reservoir to the system and monitor its temperature What are the types of temperature probes available There are 6 and 12 permanent temperature probes available as well as a flexible implantable probe Gas Monitoring System How are gas monitoring CO O2 and pH monitoring typically performed The system that HART 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 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 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 as well as individual protocols The extraluminal space of the oxygenator is typically supplied with 95 air 5 CO
28. amber 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 37 1 8 tube 1 8 rigid tube 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 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 Nov 01 2013 Revision 1 4 1 Cap mounting Plastic Bottle Large Animal Luer Lok 1 4 28 1 8 barb 3 8 Barb 1 8 NPT 1 4 NPT Nuts 38 Modified 1 2 Barb 3 8 NPT Modified 1 8 tube 1 8 NPT Modified 1 4 Barb 1 4 NPT To maintain sterility when not in use apply caps to each barb Plug Barb 4 pk 10 31112 Plug Barb 8 pk 10 31113 Plug Barb 2 pk 10 31114 Plug Barb 4 pk 10 31115 7 1 4 28 nut 1 2 Barb 1 4 NPT 3 8 NPT Nuts 1 4 Barb 1 8 NPT 1 4 NPT Nut Nov 01 2013 Revision 1 4 1 Luer lock utility port in Return from chamber inch out reservoir to left atrium 1 aorta trachea outlet Luer lock utility port out chamber or right a
29. ant Rate Constant Rate 3 Constant Rate i Pulsatile Cycle Total Run Time Temp 2284 Temp 29 39 sc Temp NaN C Temp NaN C Running Time Press NaN mmHg Ava Press 2 60 mmHg Avg Press 2 49 mmHg Avg Press 0 05 mmHg Avg Paused Time Rate 0 00 ml min Rate 0 00 ml min Rate 0 00 ml min Rate 0 00 ml min i 1second Temperatures Pressure FlowRate Pump iCharts Pump 2Charts Pump 3Charts Pump 4Charts Analog Inputs Logging Rate 190008 vyf EE Experiment Log Pump 2 Pressure Average 4 100 Time Function Pump Value amp Pump 2 Pressure Raw V 3 75 UCL 57 LCL 92 2 0 Flow Command CH 1 t 300 In the above state no data logging is occurring Clicking the green button will begin the program Incrementally adjusts flow rate of all pumps Adds an Experiment note When the trials are running the master controls will appear as above Use the red button to stop all data logging 41 Nov 01 2013 Revision 1 4 1 Starting Pump Programs The green icon starts the programs of all active pumps and begins data UCET TIAE NEUE logging Before data logging begins the user is prompted to either create a new file or append Experiment Name Experiment Mame 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 ARVARD APPARATUS engen 3 5 2013 10 38 2
30. 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 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 loc
31. ated 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 70 Nov 01 2013 Revision 1 4 1 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 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
32. ber 110V 31097 14 chamber 220V 31195 10 chamber 110V 31194 10 chamber 220V 31144 For All Chambers Set up the PID Controller for all blanket heaters in use Connect the heater power cord into the PID Control box Connect the temperature probe into the PID Box Note Refer to Appendix B for more information about PID controller theory and application 18 Nov 01 2013 Revision 1 4 1 For 5 5 Chambers a Orient the bottom heater pad with the power in cable so as to fit between the feet and drain valves Blanket Heater Assembly for 5 5 chambers 5 5 chamber 110V 31048 5 5 chamber 220V 31193 b The heater pad cord coming from the bottom should be oriented so it will come up the back of the chamber c Attach the two short springs between the metal pegs dowel pins sticking out of the bottom plate 4 d Wrap the 2nd heater pad against the back of the chamber and attach with the two long springs through the grooves in the heater e Fasten the springs by placing the spring hook into the loop 19 Nov 01 2013 Revision 1 4 1 13 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 14 The level of liquid in the chamber can be determined by cutting the rigid Teflon tubing inside the chamber to the desired height Depending on the pr
33. bnet mask should be 255 255 255 0 Power Cable 31076 U S 31077 Euro see parts list for other countries Temperature Probe 30960 12 probe amp diameter 30959 6 probe 4 amp diameter 31079 12 probe diameter 31078 6 probe s diameter Reservoir Sizes w 31105 500mL Heater B 31104 1000mL 30250 110V 31100 2000mL 30247 220V 31099 4000mL Note 1 8 diameter temperature probes use a 1 8 compression fitting while 1 16 diameter probes use a 1 16 Touhy Borst fitting 31059 13 Nov 01 2013 Revision 1 4 1 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 Communications cable to laptop and ORCA Controller Cable color may vary Note In order to conserve valuable bench space some users have found it useful to set the controller horizontally and to place the laptop on top of the controller 4 Consider which pump to designate Pump 1 Pump 2 etc The software will identify whichever pump is plugged into the top left port on the controller as Pump 1 It may be easier to identify the pumps if a label is attached to each drive motor see image at bottom right For each pump in use connect pump communication cord to the Dack of the pump and to the corresponding location on the ORCA Controller Note Some users stack pumps on top of another to save
34. ch the set delivery rate Diastole rate is the rate in ml min that the Mele Dame pump is allowed to increase to reach the set fill 0 000 0 000 rate Diastole Rate 0 000 i 0 ion Mod In Set Pressure Mode constant pressure is set perztion ars and the pump varies the flow to maintain the pressu re setti ng Please select the Start Mode you wish Run Mode Pulsatile Cycle Set Pressure Mode gt Max rate and min rate determine the maximum minimum rates at which the pump is allowed to operate PID Settings P Gain PID Setting P Gain is the proportional gain which dictates how fast the pump speed is allowed to 0 000 0 000 guess change in order to reach the set point 0 000 Diastole Rate D Gain 0 000 0 000 PID Setting I 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 Please select the Start Mode you wish Run Mode In Inverse Mode the setting for the diastole and Pulsatile Cycle InverseMode v systole cycles are reversed Otherwise the parameters are the same as in Standard Mode Systole 9 35 000 Nov 01 2013 Revision 1 4 1 In Reverse Mode the systole is operated on during the delivery phase The diastole is operated on during the fill phase Start Mode The user has the option to set a time delay before the pump starts running at the rate at which it has been set or can opt for the star
35. d 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 4 Nov 01 2013 Revision 1 4 1 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 e ov 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 Bio
36. d 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 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 77 Nov 01 2013 Revision 1 4 1 Troubleshooting Guide The DSP4000 does Insufficient or too much Disconnect power from the not display power is being applied to DSP4000 nend add the DSP4000 Verify that the voltage is 24 VDC menus and the current is at least 1 5 Amps
37. ding 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 72 Nov 01 2013 Revision 1 4 1 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 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 mi
38. dpoint 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 73 Nov 01 2013 Revision 1 4 1 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 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 an
39. e to desired value Example photo is set to 0040 Press SET to save SV temperature You will be returned to 0001 menu 4 No other settings in the 0001 menu need to be changed Use the down up buttons to select END and press SET to return to main screen 84 Nov 01 2013 Revision 1 4 1 Auto Tuning 1 Verify device is now at main screen showing thermocouple temperature in top red display and desired Set Value in lower green display 2 Press and hold gt button until AT light blinks OUT light will also be flashing at different intervals showing heater output Status se 5e re te T BEE 21 10 f 3 Auto Tuning should take several minutes depending on chamber size to be heated When tuning is complete AT light will stop blinking and remain off The PID Controller has now determined the correct heating parameters OUT light will continue to flash as the PID controller maintains the Set Value The PID Controller will save the settings after powering off so Auto Tuning should not need to be run more than once for a given configuration If desired Auto Tuning can be run again by pressing and holding the gt button Note The first time running the PID controller it is recommended that warm water is used to reduce the amount of overshoot as much as possible The PID control will test limits and will intentionally overshoot and undershoot the set point range which can damage an organ After the a
40. er 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 The camera button lets you take a snap shot and save it on the right screen while still ds displaying the left screen as live image 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 Note The Image Capture System is still in beta some functions may not yet work properly in which case other software may need to be supplemented 56 Nov 01 2013 Revision 1 4 1 Chapter 5 Care amp Maintenance 5 1 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 Stainless steel parts may be sonicated moog CAUTION Do NOT use bleach as it can cause the chamber to crack and subsequently leak Cross Contamination Prevention Biohazardous Waste and Product Disposal Cross Contamination Prevention Universal 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 p
41. es Variables Pump 1 Temperture Select Pump 1 Pressure J Select Pump 1Rate V Select Pump iRaise Time Select Pump iDescentTime Select Pump 1Period Select Pump 2Temperture Select Pump 2Pressure Select Pump 2Rate Select Pump 2Raise Time Select Pump 2Descent Time Select Pump 2Period Select HARVARD APPARA 3 5 2013 8 39 47 AM iv Window Print Temp Pumpi Pump4 JV Press Pump2 f Analoog In How Pump3 IV Email TextMessage Pump 3 Temperture Select Pump 3Pressure J Select Pump 3Rate IV Select Pump 3Raise Time Select Pump 3DescentTime Select Pump 3Period Select Location 1 02 Select Location 1 CO2 Select Location 1 PH Select Location 2 9502 Select Pump 4 Temperture Select Pump 4Pressure Select Pump 4Rate IV Select Pump 4Raise Time Select Pump 4Descent Time Select Pump 4Period Select Pulsalite Pump BPM actual Select Pulsalite Systole ms J Select Pulsalite Diastole ms Select Cyde Bool Select 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 computer s My Documents folder and can be copied onto a portable drive and taken to another PC for evaluation File Operate Tools Notification Support B x Rate 1501 Run5 gt 3 Default gt x Pump 1 Pump 2 ml min 100 00
42. f 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 096 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 69 Nov 01 2013 Revision 1 4 1 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 calibration 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
43. for the channel from DSPCALIB DAT PPB Dissolved 0 PPB 5000 PPB in the USB KEY Factor 1 for channel 3 in Figure 4 9 v Level A ree Gas Phase 0 PPM 5000 PPM Ei 180 PPB mn lt PPB 5000 PPB A Level eo Gas Phase 0 PPM 5000 PPM Nov 01 2013 Revision 1 4 1 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 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 calibration 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 99 o DO 100 Air Saturation 95 DO 454 Air Saturation I
44. hannel 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 66 Nov 01 2013 Revision 1 4 1 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 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 wh
45. heet and import it instantly into TMS It is also designed to allow you to share datasets between institutions and there s even a link to eMuseum Network that can help you work wnloadir j ually enter r collaboratively by downloading records from a lending institution now there 10 neg gt manually enter records for loan objects Analog Inputs The ORCA controller allows you to feed in any analog signal and this software can capture display and save the traces Temperatures Pressure Flow Rate Pump i Charts Pump 2 Charts Pump 3Charts Pump 4Charts Analog Inputs Analog Input 1 Pa J owm y 1 Analog Input 2 4 oos s 09 Analog Input 3 a 0 013 0 8 Analog Input 4 n 00 0 7 0 6 0 5 0 4 0 3 0 2 1 ITE 1AF nr Aet LV li ond pa 2 2 l I I l l l l l l l l I I I l l l TG A AS sj Sr See See ee dH Ge hg Rh ten du c cn Ze Is QC I Time seconds 54 Nov 01 2013 Revision 1 4 1 4 4 Heater Temperature Control amp Pressure Channel Calibration Heater Temperature Control Heater Temperature Control Access the Temperature Control menu through Tools Heater Heater Controls Probe Ta Control Probe to Control allows the user to select between four m xj different probes labeled TC 1 to TC 4 Heater Control Heater Control can be set to either Enabled or Disabled J Disabled SetPoint allows the user to set
46. 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 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 dif ferent 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 supple ment 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 outlined 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 O2 O2 Air Sat O2 PPM or PPB or SUB PPB O2 mmHg O2 deg 64 Nov 01 2013 Revision 1 4 1 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
47. i e forward flow is always designated as flow from left to right This will help avoid potential confusion 24 Nov 01 2013 Revision 1 4 1 3 3 Chamber Specific Flow Path Setup i 14 Chamber Large Animal System Example Configuration This is an example diagram representing a possible configuration for the ORCA bioreactor for a 14 chamber large animal system This is intended as a guideline to assist in setting up tubing pathways Be sure to follow individual experimental procedures Your laboratory s setup may differ from what is depicted below this diagram is not a replacement for your laboratory s protocols Peristaltic Pump 3 KEY T T fitting Z ON Single Channel Pump Head ZooN Dual Channel Pump Head O2 Heater with Reservoir inside Peristaltic Pump 2 Organ Chamber 25 Nov 01 2013 Revision 1 4 1 ii 10 Chamber Large Animal System Example Configuration This is an example diagram representing a possible configuration for the ORCA bioreactor for a 10 chamber large animal system This is intended as a guideline to assist in setting up tubing pathways Be sure to follow individual experimental procedures Your laboratory s setup may differ from what is depicted below this diagram is not a replacement for your laboratory s protocols Peristaltic Pump 3 Peristaltic Pump 1 KEY T T fitting Z ON Single Channel Pump Head ZooN Dual Channe
48. ile 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 O Ze OL E STAR Channel 1 RTD1 Black 1 RTD1 Red 2 RTD1 White 3 TM Power In 05 000 T I Process Monitor Channel 2 RTD2_Black 4 RTD2_Red 5 RTD2_White 6 DSP4000MC Channel 3 RTD3 Black 7 RTD3 Red 8 RTD3 White 9 KOCTKM TKM TKM ZEM Channel 4 RTD4 Black 10 RTD4 Red 11 RTD4 White 12 MN meme 1 manna mann ITI mu 9232 4 20 mA Inputs l Temperature Pressure 4 20 mA Outputs e Store the cylindrical plug in a safe place Step 5 Insert the bare wires as per the above table Jan 31 Clamp down using screwdriver NOTE The white wire is 4 now black the black are interchangeable SS amp Connect the threaded RTD probe tip into your sensor The Flowcells and the Tri fold Flowcells both have built in threaded ports to accommodate the RTD probe 67 Nov 01 2013 Revision 1 4 1 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
49. indow Nov 01 2013 Revision 1 4 1 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 Tubing Size and or add on Calibration Factor selected The calibration factor can also be adjusted Single Head L S 17 1 Head Configuration Select from single double triple or quadruple head configurations Note The max speed of the motor is 300rpm Tubing Size Select the PharMed tubing size that you wish to use See table below for a description of the available 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 know the tu bing size Please select Head Configuration Tubing Size and or add on Calibration Factor being utilized for each pump head and verify that nz CHEER the correct tubing size is selected The pump Pump 2 C a n n ot V e rify th j S f O r th e u S e r Please select Head Configuration Tubing Size and or add on Calibration Factor Head Configuration Tubing Size Calibration Factor Calibration Factor The number set for this parameter Flow Rates is the inverse of the variance of the liquid delivered The Mes 140 lion calibration f
50. 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 measurement channel The configuration utility is also used to reassign channels to different measurements 76 Nov 01 2013 Revision 1 4 1 8 Cleaning and Maintenance Optical Sensors are not affected by ambient light when in use however they are sensitive to light an
51. l 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 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 36 Nov 01 2013 Revision 1 4 1 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 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 1 4 rigid tube chamber circulation 1 4 tube ch
52. l Pump Head Heater with Reservoir inside 26 Nov 01 2013 Revision 1 4 1 Large Animal Heart System 10 Chamber LE Pulmonary artery Right Ventricle In from aorta m Out to Reservoir Out to peristaltic pump 27 Nov 01 2013 Revision 1 4 1 Large Animal Heart System 10 Chamber Aorta Pressure 2 Right Atrium Pressure Spare Plug Port Left Atrium Pressure 1 uem 2 e Chamber or Spare Pressure 4 Temperature Probe Chamber Vent Left Ventricle Organ elevator handle Pulmonary Ar Compression lock screw tery Right Ventricle Arm Articulator holder Aorta Feec T Additional Sensor Port 30470 Chamber Quick Connect Fill Chamber Quick Connect Drain 31211 g 31211 g 31210 Pa 31710 z d 28 Nov 01 2013 Revision 1 4 1 Large Animal Heart System 10 Chamber 29 Nov 01 2013 Revision 1 4 1 Large Animal Aortic Valve System 10 Chamber 1 2 Diastolic after load pressure valve Tighten clockwise to increase pressure 30 Nov 01 2013 Revision 1 4 1 iii 5 5 Chamber Small Animal Lung System Example Configuration This is an example diagram representing a possible configuration for the ORCA bioreactor for a small animal lung system This is intended as a guideline to assist in setting up tubing pathways Be sure to follow individual experimental procedures Your laboratory s setup may differ from what is
53. ll 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 rec ommend 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 Nov 01 2013 Revision 1 4 1 2 4 Equipment Components It is important to 1 Ensure that the ORCA Bioreactor package was received completely and without damage if the package arrived as damaged contact your local technical support group Do not use damaged parts 2 Store the ORCA Bioreactor in a cool dry place free from dust and other potential contaminants until ready to use Overview of Main Components A Heater Reservoir B Peristaltic Pumps C Pulsatile Pump D Organ Chambers E ORCA Controller F Laptop with Data Acquisition Software 8 Nov 01 2013 Revision 1 4 1 Heater Reservoir The heater is used to maintain temperature throughout the system to emulate physiological conditions Reservoirs have built in ports for interaction with the organ chamber and the oxygenation system and a special port for the thermocouple which monitors temperature within the reservoir Pumps Pumps are selected according to the o
54. 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 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 Dataq 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 pH Scale read for the channel from arameter DSPCALIB DAT in the USB KEY Factor 2 d Factor 3 for channels 2 and 4 in 0 Ommhg sooommio Figure 4 9 95 Air Sat 100 Saturation CO High scale value LET 0 PPM 5000 PPM read
55. mediately oO Stop on Time Target Ozn HARVARD APPARATUS Step Viewer Selector Step Name gate 150r Runs 0 Step Description ev Start Immeditly Run a Constant Rate 150ml min Infuse Direction Run for 5Seconds lv Constant Rate Q Start Immediately Oo Stop on Time Target 7 ik Oms 155 0 Would you like to up date the name and Description Yes No 1 1 I 1 1 1 1 1 1 1 1 1 1 1 07i 55 0 52 5 50 0 47 5 45 0 42 5 40 0 37 5 35 0 32 5 30 0 27 5 25 0 22 5 20 0 17 5 15 0 12 5 10 0 7 5 5 0 2 5 0 0 44 Do you wish to create a new step with this configuration or up date the selected step If you select up date any protocols using this step will be up date also Best practice would be to create new unless you are sure of all usages of this step Step Name Rate 100I Run5 Step Description Start Immeditly Run a Constant Rate 100ml min Infuse Direction Run for 5Seconds New Up Date cancel j po u Create or Select Step Would you like to Create a New Step with your present configuation W Create Step Name Ratei50I Run5 Select select Start Immeditly Run a Constant Rate i5 ml min Infuse Direction Run for 55econds When the step parameters have been selected a yellow gear with a green cross will pop up The user must click this gear and save the step for it to be implemented Otherwise any changes made will be discarded upon leaving the w
56. mming Instructions Setting Thermocouple Type 1 Press SET to enter password menu Bottom display will show 0000 2 Enter 0089 using arrow buttons a Use button to change selected digit down up buttons to change digit value b Press SET to enter 0089 menu 3 IntY is the first item in 0089 menu Press SET to edit IntY which is the thermocouple type setting a Default value is P100 use down up buttons to scroll to t The Bioreactor PID Temperature Controller is only compatible with type T thermocouples b Press SET to save thermocouple type You will be returned to 0089 menu 4 No other settings in the 0089 menu need to be changed Use the down up buttons to select END and press SET to return to main screen 83 gU AL1 PV Paty UL gt e Nov 01 2013 Revision 1 4 1 Inputting Temperature Set Point Value Note If the user allows the PID controller to idle for about one minute the controller will return to the main menu If this happens simply follow the above steps again to get back to the desired menu 1 Press SET to enter password menu Bottom display will show 0000 2 Enter 0001 using arrow buttons a gt button to change selected digit down up buttons to change digit value b Press SET to enter 0001 menu 3 SV is the first item in the 0001 menu Press SET to edit SV which is the Set Value a Default value is 0800 use arrow buttons to chang
57. munications cable is plugged into computer and into the ORCA controller e he ORCA controller is turned ON e Your computer s static IP should be set to 172 16 51 59 the subnet mask should be 255 255 255 0 DO NOT PLUG THE COMMUNICATIONS CABLE INTO A ETHERNET PORT AND CONNECT IT TO THE UNIVERSITY INSTITUTION NETWORK THE LAPTOP COULD CHANGE THE IP ADDRESS AND NOT BE ABLE TO COMMUNICATE CORRECTLY WITH OR CA YOU WILL THEN HAVE TO MANUALLY CHANGE THE IP ADDRESS BACK TO 172 16 51 59 the subnet mask should be 255 255 255 0 Peristaltic Pumps Problem There is an issue with the peristaltic pumps concerning the backfill siphon of the reservoir into the chamber e Watch how the pump rollers stop to ensure that they always occlude the tubing 88 Nov 01 2013 Revision 1 4 1 UPDATE LOG 13 Oct 01 Created update log Added thermocouple ordering information pg15 13 Oct 08 1 3 2 Created new example config schematic for 5 5 chamber lung system pg29 Fixed minor error in PharMed tubing specifications chart pg41 13 Oct 22 1 4 1 Updated part numbers and drawings Added in Blanket Heater instructions Updated FAQs and troubleshooting section 89 Nov 01 2013 Revision 1 4 1 HARVARD APPARATUS REGENERATIVE TECHNOLOGY Harvard Apparatus Regenerative Technology 84 October Hill Drive Holliston MA 01746 USA www HARTregen com 508 893 8999 cservice HARTregen com 90 Nov 01 2013 Revision 1 4 1
58. ns across multiple experiments The image capture software allows for images to be captured in real time from cameras monitoring visible light IR UV and fluorescence other systems such as ultrasound ECHO are supported 6 Nov 01 2013 Revision 1 4 1 Chapter 2 Getting Started 2 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 2 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 2 3 General Safety Requirements The following conditions must be met prior to using the ORCA Bioreactor 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 a
59. o maintain constant pressure e Pressure Oscillation adjusts the flow to bounce between two set pressure points e Pulsatile Cycle provides a pulsatile flow Please select the Start Mode you wish Run Mode Constant Rate __StandardMode Flow Direction In Constant Rate mode flow is set by the user and derbi maintained at a constant rate Flow direction may be 100 gt changed In Constant Pressure mode constant pressure is set by the user and the pump varies the flow to maintain the pressure setting Max rate and min rate determine the maximum minimum rates at which the pump is allowed to operate PID Setting P Gain is the proportional gain which dictates Please select the Start Mode you wish how fast the pump speed is allowed to change in order to Run Mode reach the set point Constant Pressure MN Operation Mode PID Setting I Gain is the integral value used to adjust the baseline and fine tune the rate at which the pump is nn allowed to move to reach the programmed setting Constant Pressure Average Mode T Note For more information regarding PID controllers see well Appendix B PID Settings Users can also choose between Standard Mode and the joue oon am Average setting Standard Mode uses the raw data of each shes reading A reading is taken about every 25 nanoseconds gs If the signal is very noisy the Average setting is a better option which uses a rolling average of the last 40 readings for
60. ocess by running the pump faster and faster as time goes by this is considered integral control T However making changes that are too large for a small amount of error leads to overshoot Repeated changes that are too large leads to an output that oscillates around the set point Derivative control D can be used in order to dampen these oscillations by making inferences about the future based on the slope of the error function at a given point The user has the ability to set values for these parameters Generally the proportional term P should constitute the majority of the output change A small P value results in a small output response to a large input error In other words the controller will take a long time to offset the error but generally the system will be stable In contrast a large P value will offset any errors quickly but at the risk of system instability The integral term is useful for accelerating the process towards set point and for eliminating residual steady state error that could occur using a pure proportional controller However it can cause the present value to overshoot the set point value The derivative term is currently not used with the ORCA due to its inherent sensitivity to measurement noise A large sudden change in measured pressure could cause erratic changes in the control mechanism that degrade performance In physiological systems it is generally more important to minimize overshoot than to have a
61. otocol the user may want to cut the drain tube at an angle with the low end of the cut toward the chamber wall then file the point so that it is not sharp Note If the user wishes to set up a complete flow path without having to sacrifice an organ then a piece of tubing may be added in the place indicated see above right image to test the unit without an organ in place Note The polycarbonate chamber windows shown below for the 5 5 10 and 14 chamber respectively are durable but can be prone to getting scratched Tf the windows become too scratched then replacements may need to be ordered The replacement part numbers are shown below is 5 5 31083 includes gasket 10 31084 pra 31085 includes gasket includes gasket 20 Nov 01 2013 Revision 1 4 1 15 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 For each size of tubing cut a 1 piece and glue laterally as well as a Ya piece glued so that the internal ID of the tubing is easily displayed This is useful because otherwise it is often difficult to tell the correct size tubing needed at a glance See page 22 for a reference guide 16 Any barb fittings that are not in use should be sealed with the appropriately sized caps in order to maintain the system s sterility There are 3 8 1 2 and 3 4 caps available They are sho
62. ovide flexibility in flow range as well as the number of channels required for the most challenging protocols a eae PE y M ENIMS Ki TN One Single channel Two Single Four Single channel Head channel Heads Heads 31061 31062 31063 One Dual channel Two Dual channel One each Single and Head Heads 31065 Dual channel Heads 31064 131066 Additional Part Numbers 31067 Mounting Screw for Two pump heads 31068 Mounting Screw for Three pump heads 31069 Mounting Screw for Four pump heads Pulsatile Pumps 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 Nov 01 2013 Revision 1 4 1 Chambers Solid Organ Chambers Large Solid Organ Chamber 14 30240 holds approx 32 8L Intermediate Solid Organ Small Solid Organ Chamber 10 30260 Chamber 5 5 30320 holds approx 11 6L holds approx 2L 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 recellularization procedures Chamber access ports are built in for removal and addition of media Numerous sam
63. pling ports are included and the built in windows allow for manual access to an organ Hollow Organ Chambers 30300 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 See the Hollow Organ Bioreactor manual for additional information 11 Nov 01 2013 Revision 1 4 1 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 31072 Pump Drive with appropriate heads 1 to 4 The type and number of heads will vary with each system 4 channel 3 2 S controller 31082 E T cable 31073 Laptop and power cables will vary depending on the order 12 Nov 01 2013 Revision 1 4 1 Communications cable 32077 DO NOT PLUG THE COMMUNICATIONS CABLE INTO A ETHERNET PLUG AND CONNECT IT TO THE NETWORK THE LAPTOP COULD CHANGE THE IP ADDRESS AND NOT BE ABLE TO COMMUNICATE YOU WILL THEN HAVE TO MANUALLY CHANGE THE IP ADDRESS BACK TO 172 16 51 59 the su
64. ratus 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 3 General Description Application 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 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 63 Nov 01 2013 Revision 1 4 1 though the total sensor capacity may differ In the descriptions below all configurations will be re ferred 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 for which the connected sensor s is are configured pH O2 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
65. rgan 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 and pulsatile 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 pumps and a pulsatile blood pump 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 pumps may be added at any time Additional peristaltic pumps add flexibility to accommodate elaborate protocols but are not required 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 Nov 01 2013 Revision 1 4 1 Pumps Peristaltic Pumps Peristaltic pump heads are available in single and dual channel versions High Capacity up to 1200mL min head Mid Capacity up to 500mL min head Up to four 4 heads may be used on each peristaltic pump to pr
66. rotect 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 57 Nov 01 2013 Revision 1 4 1 5 2 Replacement Parts List Part Part Number Barb Plug 1 4 pk 25 Fitting Horizontal Chamber Drain Level 25ea ting Luer Lok x 1 16 NPT 25 a 31113 Fitting Right Angle 14 Chamber 25ea 30420 Barb Plug 3 4 pk 10 31114 Fitting Luer Female 1 4 28 25ea 31163 31097 Fitting 1 4 Polycarb Straight w Luer Port 25ea 30904 31144 Fitting 3 8 Polycarb Straight w Luer Port 25ea 30899 31048 Fitting 1 2 Polycarb Straight w Luer Port 25ea 30905 30509 Fitting Tee 25 ea 31138 30564 Fitting Tee 25 ea slip Luer 512294 Bubble Trap 1 2 Be Heater Reservoir 115V 30250 Bubble Trap 1 4 30450 Bubble Trap 14 head 30295 Heater Reservoir 220V d Bubble Trap 10 head 30442 Liner for Heater 25 each 30298 aa SE 30456 Nut Luer Clear 25 each 31141 32007 Nut Luer Green 25 each
67. ry 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 HART 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 the researcher and the physicians to accelerate the research and utilization of that research in patients 5 Nov 01 2013 Revision 1 4 1 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 conditio
68. s 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 nke Step Mode See Step Mode for additional info Start Mode See Start Mode for additional info 44m Stop Mode See Stop Mode for additional info Repeat Mode See Repeat Mode for additional info Pump Selector ta Choose from peristaltic syringe pulsed blood or ventilator modes Nov 01 2013 Revision 1 4 1 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 Name gate 150r Runs T Step Description dia Start Immeditly Run a Constant Rate 150ml min Infuse Direction Run for 5Seconds op Constant Rate O Start Immediately oO Stop on Time Target Repeat Increase n 1 1 1 1 1 1 1 1 1 zn I 55 0 52 5 50 0 47 5 45 0 42 5 40 0 37 5 35 0 32 5 30 0 27 5 25 0 22 5 20 0 17 5 15 0 12 5 10 0 7 5 5 0 2 5 0 0 HARVARD APPARATUS Step Viewer Selector Step Name gate 1501 Runs si m ep Description odia Start Immeditly Run a Constant Rate 150ml min Infuse Direction Run for 5Seconds De Constant Rate Q Start Im
69. s w Luer Port for Pressure Transducers 1 4 30904 3 8 30899 1 2 30905 16 Nov 01 2013 Revision 1 4 1 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 swabable port This provides an insulating fluid between the transducer and the medium without a negative effect on the pressure readings 11 If not already assembled 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 Swabable ports 31081 pk 25 ONLY AUTOCLAVE ONCE to maintain integrity Bubble Trap Head 14 chamber 30297 10 chamber 30450 17 Nov 01 2013 Revision 1 4 1 12 Attach the blanket heater to the chamber For 10 amp 14 Chambers a Place the heater pad on the back of the chamber b Connect the springs to the hooks Blanket Heater Assembly for 10 amp 14 chambers 14 cham
70. t 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 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 recor
71. t immediately setting Ramp allows the user to select the speed at which the pump will deliver There are options in increments ranging from very slow to very fast None is also an option for type of ramping Note that the maximum flow rate is a constraint set by the type of tubing used 49 Please select the Start Mode you wish Run Mode Pulsatile Cyce ReveeMode Start Mode Please select the Start Mode you wish StartMode Start Immediately Start Mode Please select the Start Mode vou wish StartMode Ramp Mone Ultra Fast Very Fast Fast Medium alm Very Slow Ultra Slow Start Immediately Please select the Start Mode you wish StartMode Start After Time Delay Time Nov 01 2013 Revision 1 4 1 Repeat Mode Repeat Mode Please select the Start Mode you wish Four modes of repeat are allowed RepeatMode No Repeat Repeat Till User Stop Repeat Number of Times Repeat Increase n Times e No repeat e Repeat until e Repeat of times e Repeat increase N In 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 ee 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
72. there is no limit Please select the Start Mode you wish RepeatMode Repeat Number of The Flip Direction switch up allows the flow to be reversed M on every other cycle 10 repeats with flip direction switch t up would result in half of the cycles 5 in the forward direction and half 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 Count allows you to set the number of iterations EL RepeatMode Change by allows you to set how much to increase the NOR UNUS Running speed of the pump for each time the step is repeated 5 4 Max value is the maximum flow in ml min this is limited by the tubing o 0 Keep Pump Running allows the pump to continue to run as the final flow rate is reached x 50 Nov 01 2013 Revision 1 4 1 Notifications This window allows the user to enter contact information in order to be notified by email when certain events 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 Saving Files Notifications User List HARVARD APPARATU 3 5 2013 8 38 29 AM email Address jbouchard harvardapparatus com Telephone Notifications System Status User Jeff Bouchard Y Iv Enable Internval _15minut
73. tion How is temperature regulation achieved The reservoir heater is a major source of heat and temperature maintenance for the system but it not sufficient as a standalone All chambers use at least one wraparound heater in conjunction with a PID system in order to maintain physiological temperatures 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 denatured If medium from a cold room 4 C is introduced the time required to raise the temperature to 37 C will be very lengthy The reservoir bottles come in various sizes and with different ports to accommodate a wide variety of protocols How is temperature monitored The ORCA controller provides the ability to use four temperature probes A temperature probe is normally placed in the reservoir to control heating functions and heat the medium when the temperature dips below the set point with a maximum of 39 5 C Another probe is used to monitor the temperature within the chamber itself An implantable temperature probe can also be used to measure temperature inside an organ Addition
74. 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 62 Nov 01 2013 Revision 1 4 1 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 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 Appa
75. trium 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 39 Nov 01 2013 Revision 1 4 1 4 3 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 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 In 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 Be sure of the following before running the software 1 Windows auto updates is turned off otherwise it could cause the computer to restart whilst running an e
76. uto tune function has been used once it is ready to use with an organ 85 Nov 01 2013 Revision 1 4 1 Appendix C Frequently Asked Questions amp Troubleshooting ORCA Bioreactor Chambers What are the sizes of the chambers available There are three different sizes of chambers available 14 10 and 5 5 in diameter The right selection is simply based off of the size of the organ being studied How are the chambers sterilizable Chambers and their components are designed to be sterilized by normal laboratory methods such as autoclaving EtO and plasma sterilization How are the chamber lids designed Multiple ports are available in the top lid of the chamber to accommodate the various cannulation and access requirements For what range of pressures are the chambers designed The chambers are designed to hold physiological pressures They are NOT designed to be used as pressure chambers which allow 1atm 15psi to be applied The chambers can handle negative ventilation at several psi Pumps What are the maximum minimum flow rates achievable by the peristaltic pumps The peristaltic pump drive units are very flexible in their design and can have up to four heads on each drive unit Each of the heads can be either single or dual channel The single channel heads allow a flow rate of 1 2L min Therefore the maximum rate per pump drive unit is 4 8L min by using 4 pump heads This is sufficient for the majority of applications In
77. wn below 1 4 Plug pk 10 31112 3 8 Plug pk 10 31113 1 2 Plug pk 10 31114 3 4 Plug pk 10 31115 21 Nov 01 2013 Revision 1 4 1 Pump Head Config Single Channel Head Two Single Channel Heads combined by a Y barb Four Single Channel Heads combined by a Y barb Two Dual Channel Heads 4 channel Tubing 1 32 0 8 mm 32 156 mm 1 16 1 6 mm 4g 188 mm 1 8 3 2mm l4 25 mm 1 8 318 mm 3 16 4 8mm g 313 mm 1 4 6 4mm 3 3 375 mm 1 4 6 4mm 5 16 7 9mm he 11 1 mm 3 3 9 5mm 1 2 12 7mm 11 12 7mm 3 4 19 1mm 22 8 e ec gt Typical Function s e Perfusion into organ for smooth high flow rate e Empty chamber to reservoir e Highest flow rate Flow to oxygenator Flow to pre organ O sensor Flow to post organ 02 CO pH sensors Flow from a medium container e May be added to the pump with two dual channel heads to allow addition and removal of reservoir medium e Maximum flow rate 1 2 l min total of 4 8 l min Teflon Translucent more Tygon Clear flexible L S 13 31101 L S 14 31102 A L S 16 B L S 16 30340 30341 are er L S 25 30339 L S 17 L S 17 30338 30342 L S 18 30337 30552 30344 P J 30553 Nov 01 2013 Revision 1 4 1 23 173 connector 17 connector 3 8 connector M eme om aon 1 7 connector
78. xperiment 2 The ORCA power cord is plugged into the ORCA controller and the wall 3 The Communications cable is plugged into computer and into the ORCA controller 4 The ORCA controller is turned ON To run the program select the Bio 1 initiation icon from the Desktop When finished with an experiment it is important to shut down the ORCA controller software using CTRL Q or the File gt 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 40 Nov 01 2013 Revision 1 4 1 Improper Shut Down may corrupt the config file If the config file is corrupted a back up is available to correct the situation 1 Open My Documents folder and delete the Harvard Data folder 2 Copy the BackUp Harvard Data folder into my Documents 3 Rename the BackUp Harvard Data to Harvard Data The Main Screen gt BIO 1 m 5 x Fie Operate Tools Notification Support Bee d 9 i pen HARVARD APPARATUS Pump 1 Pump 2 E 3 5 2013 10 28 43 AM p AP AP oe Experiment Name Experiment_Name Status o Oo Oo Experiment Time Data Start Time ml min 7 100 00 mi min 100 00 ml min 40 00 ml min Const

ORCA ™ Bioreactor Manual

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