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Solid Organ Bioreactor Manual - Harvard Apparatus Regenerative

1. Manual EY d Device 1 System Logging Confiq System Logging Configuration Logging Location System Logging Config System Logging Configuration Logging Location E le Logging Rate Fastest 0 05 sec x y Fastest 0 05 sec 0 5 seconds 1 second A Sal Configuration Logging Rate allows frequency of data collec 10 seconds tion points Collecting 4 channels of data every minute will result 15 seconds in a data file of 1 Meg per day 30 seconds 1 minute Revision 2 8 75 July 15 2015 ERST Regen al egen vi File Apps Pumps Heaters System Notification Support Manual H RT i 10 8 2014 12 32 15 PM A Regen 3 Device 2 Event Log Manual Device 1 Device 3 Device 4 12 0712 PM Device 1Step Changed 12 07 15 PM Start Device 9 Rate of 100 00 ml min 12 07 40 PM Stop Device 1 12 07 58 PM Start Device 1 Rate of 100 00 ml min 12 08 30 PM Stop Device 1 12 09 05 PM Device Repeat Mode Changed 12 09 08 PM Device 1Parameter Changed 12 09 10 PM Device 1Stop Mode Changed 12 09 29 PM Start Device Rate of 100 00 ml min 12 10 05 PM Stop Device1 12 10 07 PM Start Device Rate of 100 00 ml min 12 10 11 PM Stop Devicel m n C LLA es S a E d ig o es e e e a x gt A wv e UIP VX L 100 0 ml min Mode Constant Rate 12 00 ml min Mode Constant Rate
2. Tama HHM foom amp i Temperatures Pressures Ficw Rates Device Charts Devel Chats Device 3 Charts Deoced Charts Analoglnputi Userd User2 Uses User Jo Tempentwel Jo Temperatwel To Tempar L Temperature 4 s doneert tern F TO blankettemp Phot Add Pict Eraut analog uut 1 Apply Pt Remove Pict Defaut Pets Change auda tale eth room dens Page chart to pan iroaM mijam dondom iodsam iad as 10 14 20 AM 101423 444 Changs y inds state by dovble chcking on min maxvaloes Tere Hit MSS Zoom Q en beiang 3 Select the CHART Tab Layout Single Chart 4 Under PLOT select Add a plot Tab Name Temperstures oF Select Apply Pot Chart Top Chart Plot Add Plot Input Analog Input 3 Apply Plot RemovePlot Default Plots Change x axis scale with room slider Pause chart to pan Change y axis scale by double chclong on man max values Revision 2 8 104 6 Select the calibration tab 7 Select the appropriate Anal g input that this sensor is connected to 8 Select The zero Span 9 Press the index button to get to the set point 1 screen Revision 2 8 PUE Calibration Ingut Analog Input 3 X Type Select Select Input and Calibration Type above Calculated Measured Value Raw Signal Chart 104545 11 5 11 LJ c tr a 1 Amplitude E kn i Calibration Input Analog Input 3 pe
3. 200 J Pressure 2 AN 150 E E 100 v a UCL 150 g 30 a LCL 50 i Show Limits V ue 1 10 18 PM 1 10 19 PM 850 J Flow 2 N E gt 600 E c 400 UCL 800 E TM LCL 200 AR 150 Show Limits 7 1 10 18 PM 1 10 19 PM Revision 2 8 e v ib A y 200 0 ml min Er 150 0 ml min Temperature e Pressure Pressure Temperature Device 1 Charts Device 2 Charts Jpevice3 Charts Device 4 Charts Analog Inputs Useri User2 User3 User4 I 1 10 20 PM 1 10 21 PM Time HH MM SS I 1 10 20 PM 1 10 21 PM Time HH MM SS 79 12 44 16 PM 12 44 24 PM 12 44 24 PM 12 44 33 PM 12 44 42 PM 12 44 44 PM 12 44 48 PM 12 44 54 PM 12 44 59 PM 12 45 24 PM 12 45 31 PM 12 45 32 PM Start Device 2 Rate of 12 00 ml min Device 2Details Changed Device 2Parameter Changed Device 2 Rate of 200 00 ml min Start Device 3 9 Rate of 12 00 ml min Start Device 4 9 Rate of 12 00 ml min Device 3 Rate of 150 00 ml min Device 4Parameter Changed Device 4 9 Rate of 300 00 ml min Stop Device4 Stop Device 3 Stop Device 2 System Status BEEN I 1 10 22 PM LU 1 10 22 PM 1 1 10 23 PM 1 10 23 PM 1 10 23 PM 1 10 23 PM Zoom Q IR July 15 2015 ES HART Regen File Apps Pumps Heaters System Notification Support 12 44 16 PM Start Device 2 Rate of 12 00 ml min 12 44 24 PM Device 2Details Changed 12 44 24
4. e E o 300 0 ml min Mode Constant Rate pos 150 0 ml min Run User Stop 200 0 ml min 1 of 5 Run User Stop 100 0 ml min Mode Constant Rate es E i a o o C Flow Rate 0 000 ml min Pressure 3 8 mm Hg Temperature 469 C Flow Rate 150 0 ml min Pressure 4 2 mm Hg Temperature 21 2 C Flow Rate 200 0 ml min Pressure 3 2 mm Hg Temperature 221 C Flow Rate 0 000 ml min Pressure 1 6 mm Hg Temperature 221 C 4 mil Temperatures Device 1 Charts Device 2 Charts Device 3 Charts Device 4 Charts Analog Inputs Userl User2 User3 User4 T v Flow 2 v Flow 3 v UCL 1500 Saye LCL 100 2 Nn 320 1 1 I 1 1 I I 1 1 1 I 1 1 Li Show Limits 12 44 29 PM 12 44 36 PM 12 44 40 PM 12 44 44 PM 12 44 48 PM 12 44 52 PM 12 44 56 PM 12 45 00 PM 12 45 04 PM 12 45 08 PM 12 45 12 PM 12 45 16 PM 12 45 20 PM 12 45 24 PM 12 45 28 PM Time HH MM SS a DM Flow rates Tab can be used to display any of the 4 programmed flow rates by clicking on the check box next to the Flow 1 2 3 or 4 boxes If the flows are identical they may overlap and be difficult to tell apart UCL upper control limit and LCL lower control limit refer to statistical controls that the user develops when a method is validated They are able to give you a graphic representation so
5. 8 E rx E la p E ww Uude Jj f g L o E kh o d x LL n imd ha ee i RNR RR a 1 Power on the Unit by the switch on the side of the unit in the power module This will display the home screen If the temperature probe is not plugged into the side the upper temperature read ing may be very strange as is shown 2 Plugging the temperature probe into the side will cause the upper reading to come to a normal number and will light the red SP1 in the upper left a et side A Revision 2 8 102 July 15 2015 3 Switch from Fahrenheit to Celsius Press and hold the Up and Enter button simultaneously for 5 seconds 4 After 5 seconds this screen will appear release the but tons 5 Press the index button until you scroll the menu op tions and arrive at the Unit window Ul Press the down arrow once and C will replace the F 6 Press the arrow up button and the F will replace the C 7 Press the enter button and the selection will be saved 8 To return to the HOME SCREEN either wait 45 seconds E or continue to press the index button until the Home screen appears Revision 2 8 103 July 15 2015 Calibration 1 Open the temperature tab 2 Press the gear symbol on the top right 3 i LI i LI a i 1 LI k LI i a we 191307 at 101 12 44 101315 A A 101321 A LO IT AT 103136 AM 1563339 AE LAM ZONES AR 101348 AME ML
6. APPARATUS REGENERATIVE TECHNOLOGY Harvard Apparatus Regenerative Technology 84 October Hill Road S711 Holliston MA 01746 1371 USA www HARTregen com 774 233 7300 cserviceoHARTregen com Contents Disclaimer o Overview O O 4 NEM el 6 6 6 H 13 2 23 25 30 34 38 Dl 10 1 45 a7 48 50 51 52 52 53 53 61 64 73 74 75 76 82 83 84 87 en N Revision 2 8 July 15 2015 OO Pulsatile Pump parts information Chapter 5 Care amp Maintenance 5 1 Ordering I nformation Replacement Part Numbers 5 2 Module Ordering I nformation 5 3 Cleaning co co IA co N Appendix A PI D Controller Theory amp Application Appendix B FAQs amp Troubleshooting EL EL Specifications 114 Update Log 115 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 contents or product This product is for RESEARCH USE ONLY and NOT FOR HUMAN USE Copyright 20
7. Logging Begins logging of data and allows access to logging choices 45 Revision 2 8 July 15 2015 Pump Calibration Manual auc zs 2 Press on the Pump u 2 Device 2 Please select Head Configuration Tubing Size and or add on Calibration Factor Head I mage to A choose the tubing Cn EM mm m Flow Rates selection m Kn 0 000 ml min E Pump Calibration z Mode Constant Rate Pump Calibration Pump 1 Calibration Flow Rate 0 000 ml min Please select Head Configuration Tubing Size and or add on Calibration Pump 3 Press ure 409 2 mm Hg Factor Please select Head Configuration Tubing Size and or add on Calibration Factor laN C Am Tut Head Configuration l ubing Size Calibration Factor erem e Head Configuration Tubing Size Calibration Factor Single Head L 518 l Single Head 1 T Flow Rates Mi O S min Tubing Size Size id 7 9 mm 0 31 in Max 510 ml min If no temperature probe Min 11 4 ml min Max 1140 ml min is connected the temp can read NaN Cae E Pump Calibration Head Configuration Select from single double triple or quadruple head Pump 1 Calibration configurations Note The max speed of the Please select Head Configuration Tubing Size and or add on Calibration motor is 300 rpm Factor i Head Confiauration Tubing Size Calibration Factor It is important to select the correct number g Single Head S 18 1 s of pump heads
8. Mode Constant Rate Mode Constant Rate 12 00 ml min 12 00 ml min AOLE HE l t Flow Rate 0 000 mi min Pressure 4 2 mmHg Temperature 20 9 C Flow Rate 0 000 ml min Pressure 3 0 mmHg Temperature 21 5 C Flow Rate 0 000 mi min Pressure 1 6 mmHg Temperature 21 5 C Flow Rate 0 000 ml min Pressure 3 9 mmHg Temperature 32 0 C pm 1 Pressures Flow Rates Devicel Charts Device2 Charts Device3 Charts Device4 Charts Analog Inputs Userl User2 User3 User4 l V Temperature 1 V Temperature 2 lv Temperature 3 WD V Temperature 4 Temperature C PN un UCL 40 do LCL 35 et a 20 Tu 1 Lu 1 1 1 1 I 1 1 I 1 1 1 I I Show Limits A 12 31 59PM 12 32 01 PM 12 32 02 PM 12 3203PM 12 32 04 PM 12 32 05 PM 12 32 06 PM 12 32 07 PM 12 32 08 PM 12 32 09 PM 12 32 10 PM 123211PM 12 32 12 PM 123213PM 12 32 14 PM 12 32 14 PM Time HH MM SS Zoom Q 1H Temperature Tab can be used to display any of the 4 temperature sensor readings by clicking on the check box next to the Temperature 1 2 3 or 4 boxes If the temperatures are identical they may overlap and be difficult to tell apart see above UCL upper control limit and LCL lower control limit 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 d
9. Stop on Time Target allows the operator AAO EDE AE to set a time at which the pump will automatically stop 10 Second mm Revision 2 8 58 July 15 2015 Manual Repeat Mode allows the operator to repeat an operation Device 1 automatically pressing the start button internally Four modes of repeat are allowed Please select the Repeat Mode you wish No repeat NUS NoRepeat 4 Repeat until Repeat Till User Stop Repeat Number of Times Repeat Increase n Times 4 Repeat of times 0 6300 ml min 4 Repeat increase N Mode Constant Rate CI Cancel Flow Rate 0 000 ml min Pressure NaN mmHg Temperature NaN C Please select the Repeat Mode you wish RepeatMode No Repeat In No Repeat mode the step does not repeat i In Repeat Until User Stop Mode the step will Please select the Repeat Mode you wich repeat until the user stops the program manually RepeatMode The Flip Direction switch allows the flow to be Repeat Ta Uses Stop Fis reversed Direction J o Conce Revision 2 8 59 July 15 2015 Please select the Repeat Mode you wish RepeatMode Repeat Number of Count 3 In Repeat Increase Mode the speed of the pump changes with every subsequent it zug in Repeat Number of 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
10. UCL 200 LCL 100 20 Show Limits P d Lu 1 06 22PM 10632PM 1 06 40 PM pumps i i 1 06 48 PM 1 06 56 PM I 1 06 48 PM _ 1 06 56 PM e Flow Rate 0 000 ml min 4 5 mm Hg 20 4 C Pressure Pressure Temperature Temperature 358 C Device 2 Charts Device3 Charts Device4 Charts Analog Inputs i i 1 07 04 PM 1 07 12 PM Time HH MM 55 i 1 07 04 PM 1 07 12 PM Time HH MM SS Flow Rate 0 000 ml min 4 3 mmHg i i i 1 07 20PM 1 07 28 PM 1 07 36 PM l 1 07 20 PM 1 07 28PM 1 07 36 PM 12 45 24 PM 12 45 31 PM 12 45 32 PM Start Device 2 9 Rate of 12 00 ml min Device 2Details Changed Device 2Parameter Changed Device 2 Rate of 200 00 ml min Start Device 3 Rate of 12 00 ml min Start Device 4 Rate of 12 00 ml min Device 3 Rate of 150 00 ml min Device 4Parameter Changed Device 4 9 Rate of 300 00 ml min Stop Device 4 Stop Device 3 Stop Device 2 System Status ee i 1 07 44 PM 1 07 44 PM LU 1 07 52 PM 1 1 i 1 08 00 PM 1 08 16 PM 1 08 22 PM ART 10 8 2014 1 10 23 PM H Regen Event Log Device 1 Device 2 Device 3 Device 4 v lt a Mode Constant Rate p 100 0 ml min Mode Constant Rate Flow Rate 0 000 ml min 3 3 mm Hg 21 4 C Flow Rate 0 000 ml min 2 0 mm Hg 24 6 C Pressure Pressure Temperature Temperature Temperatures Pressures Flow Rates
11. 32 4 0 mm 1 16 1 6 mm 7 16 4 8 mm AS 1 16 1 6 mm 8 3 2 mm 1030358 C ES Leno r sone 113 3 2mm 7 4 6 4 mm A 7 16 4 8 mm 16 7 9mm U eden 1 4 6 4 mm 7 8 9 5 mm D reder 1030342 X 2030346 5 16 7 9 mm 7 16 11 1 mm G B H 1030343 3 8 9 5 mm 12 7 mm 3 8 9 5 mm s 15 9 mm lj 3 4 19 1 mm 1 25 4 mm 1030553 K aon Revision 2 8 21 July 15 2015 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 it is not exact As previously mentioned it is recommended that tubes of the sizes that your lab commonly uses are glued to a plastic clip board allowing for even simpler comparison PharMed Tubing L S 13 L S 14 L S 16 L S 25 L S 17 L S 18 L S 82 Front View O QO 0O ee Tubing size Lateral View 0 15625 4 0mm OD 0 03 0 8mm ID 0 1875 4 8mm OD 0 06 1 6mm ID 0 25 6 4mm OD 0 12 3 1mm ID 0 3125 7 9mm OD 0 19 4 8mm ID 0 375 9 5mm OD 0 25 6 4mm ID 0 4375 11 1mm OD 0 31 7 9mm ID 0 75 19 1mm OD 0 5 12 7mm ID AUAU Used for connections not for use in the peristaltic pump Note When feeding the tubes through a
12. e gt Flow Rate 0 000 ml min Flow Rate 100 0 ml min Pressure 435 7 mm Hg Pressure NaN mm Hg Temperature NaN C Temperature NaN C M gen Operation Mode Selector is used to create what operations are required and how they will be implemented On this screen Run Mode Flow Rate and Flow Direction can be controlled Device 1 Run Mode There are four options to choose from Constant Rate provides a constant perfusion rate 4 Constant Pressure pump alters its flow to maintain constant pressure 0 6300 ml min Mode Constant Rate 23 4 Pressure Oscillation adjusts the flow to bounce between two set pressure points Flow Rate 0 000 ml min Pressure NaN mmHg Temperature NaN C Pulsatile Cycle provides a pulsatile flow Revision 2 8 51 July 15 2015 CONSTANT FLOW l Please select the Operation Mode you wish In Constant Rate mode flow is set by the user and Run Mode maintained at a constant rate This is regardless of the Constant Rate Standard Mode xi pressure Care should be taken not to choose a flow that is too high and can cause an IS Sue W ith th e Please select the Operation Mode you wish Run Mode Flow Flow Rate Direction ml min 100 Er organ by Constant Rate Mode increasing the Kemari TTA pressure to an Bras cm C 100 unacceptable limit CONSTANT PRESSURE RATE In Constant
13. 2015 Pulsatile Pump Parts Information Pulsatile Pump Grease 1031164 PolySulfone Autoclavable Pulsatile Pump Head 1031150 Head Removal Tool is purchased from Harvard Biosciences 5012006 1031156 1031153 1031157 1031162 1031158 1031151 1031160 p 4 i 1031154 Revision 2 8 88 July 15 2015 Chapter 5 Care amp Maintenance 5 1 Replacement Components ORCA Controller 4 1032090 9 1032092 9 ORCA Controller 34 1 1032091119 10320932 1030250 10302472 MEE 1 Dual Channel Pump Head 2 Dual Channel Pump Head Medium Reservoir Heater 1031061 1031062 1031063 1031064 1031065 3 lll 1031064 1031065 1031072 Single Channel Pump Head ual Channel Pump Head Cable 3m Pump to ORCA D PID Temperature Controller BNC Cable for PID Controller ORCA Manual Large Animal Tubing Set 1030875 1032364 1030752 1031060 Small Animal Tubing Set 1031370 8 Amp Fuse Revision 2 8 1032708 cr lt TEE TTT TY TY e July 15 2015 Chapter 5 Care amp Maintenance 5 2 Replacement Parts Descriptions O 99 Complete Reservoirs 14 Chamber 1031240 am tle 10 Chamber 1031250 BA 5 5 Chamber 1031260 14 Chamber 1 1031097 103119577 1 103104872 103119377 a NEN NEN NEN E m NEN 0 Chamber D CE nn EN Reservoir Bottles 7 EN mn a Reservoir jack O Doo y O Revision 2 8 90 July 15 2015 Cha
14. 7 16 11 1mm OD Le lt Y fittin Y 3 8 bii v Y fitting x 1 4 Y barb T fitting AV 3 8 T barb Z ON Single Channel Pump Head Zoon Dual Channel Pump Head Revision 2 8 Peristaltic Pump 2 Peristaltic Pump 3 Monitor Heater with Reservoir inside Organ Chamber Note In this setup the organ is being cannulated in two locations The tubing pathway shown through peristaltic pump 4 is optional but the user should be aware that some of the media will drip through the organ at collect at the bottom of the chamber In order to keep these levels from getting too high this pathway and pump may need to be utilized The media is pumped back into the reservoir via the Quick Connect outlet ports located on the bottom of the chamber It is also possible to cannulate the organ in a single location in which case the tubing pathway associated with peristaltic pump 2 may become unnecessary The 5 16 ID tubing will fit snugly on the 3 8 barb fittings The 5 16 tubing is used with the system because it is the largest tubing that will comfortably fit through the peristaltic pumps 25 July 15 2015 14 Chamber Lid The following is an outline of potential usages of the ports on the chamber lid and reservoir The system is designed to accommodate a wide variety of protocols Each user s setup will vary depending on
15. If the single head is select Double Heads ed the pump motor will drive the pump iple Hea low Rates A SE Tubing Size Size 75 Gain WEL head for the entire flow rate selected Nin 11 4 ml min Max 1140 ml min As an example if 100m ml min is selected as a flow rate and a single head is chosen Cancel the pump motor will turn the drive shaft at a rate that produces 100 mL min regard less if there is 1 or multiple heads mount ed If you have selected 4 quad heads and 100 ml min The drive shaft will turn at a rate that pro duces 25 mL min per head The ORCA pump is set up to produce a flow based on a single channel head There are also dual chan nel heads available that can be very convenient The ORCA pump does not know how to recognize dual channel heads Therefore it will assume that there is only a single channel head when calculating the speed of the drive If you choose 100 mL min and have a dual channel head each channel will produce 100 mL min The rate limiting factor is normally the size of the tubing as the larger tubing cannot physically fit into a dual channel head The calibration is inversely proportional to the results If you program 100 ml min and the pump is delivering 90 then a place to start is entering a value of 1 1 which should get you to 99 ml min and then refine from there Revision 2 8 46 July 15 2015 Six Standard Tubing Sizes are supported See Tubing Size Select the PharMed tubing siz
16. Iie Je 1 8 3 3 1 lie lie h16 T Reducer NO e e e e N N N N NINI A lolol e e Fe OI o Ul Ul T Reducer Luer 14 Luer 1030904 1030899 1030905 3 3 Luer 17 Luer 1032068 1032050 1032751 1031137 1032753 1032742 1032744 1032081 1032746 lg Hag 3 16 1 16 Jye lie W oo I HI P Y a s 1e 16 lie Hg a tla TJ 2 2 8 le md Y Barb Reducer W Revision 2 8 95 July 15 2015 COUPLERS D300 D150 105231 1032736 4 Inlet Outlet Media m 1032772 Fittings Inlet Outlet Gas Fittings See Female Luer to Barb Fittings of appropriate size Gas Regulator 1032565 Stand Adapter for 1032560 gas regulator Ill Il al LL I utl Ini i HEEN T Revision 2 8 96 July 15 2015 5 3 Cleaning Standard Laboratory protocols may be used In general we recommend the following steps prior to auto claving A Flushing with deionized water B Washing with a mild detergent C Flushing with deionized water Stainless steel parts may be sonicated CAUTI ON 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 b
17. Service Cservice hartregen 5555555555 txt att net Notifications System Status Status defines parameters for each user to receive notifications at a predefined interval Also al User C Senate 5 indie nar Hf Tema as Tedere P low enabling of alerts to be sent se to the text message email ad Vanables PurnpiTempersture Select Pump2 Temperature Select Pump3 Temperature Select PumpATempersture Select dress PumplPressure Select Pump2Presure Select PumpiPressum Select Pumpd Pressure Select PumplRate Select Pump2Rate Select Pump3iRate Select Pump d Rate Select Meek eat ar de M ue EE SON It is recommended that only pa User iisbiel select Uver Variabel Slet Uer Varabie3 Sit User Variable d Suit rameters to the protocol be Pulsatile Pump BPM actual Select Pulsatile Systole mz Select Pulsatile Diastole ms Select Cycle Time Select checked or the number of notifi cations may be large enough to L fatigue the user NOTI CE Three is a bug in the version 1 software that only allows notifications to be sent at either 15 or 30 minutes Revision 2 8 72 July 15 2015 M 7 Enabled H Select an Input lal Enabled Select aniaput Enable PITT TEN Select an Input Enabled Select an Input Enabled Select an Input l Enabled Select an Input Bi Enabled a Rising Falling Rising Falling 9 Rising Fa
18. for the diastole period of the duty cycle The Max Rate is the highest medium flow rate the system will allow to obtain the pressure target at any point The Min Rate is the lowest medium flow rate the system will allow to obtain the pressure target at any point PI D Setting P Gain is the proportional gain which dictates how fast the pump speed is allowed to change in order to reach the set point PI D Setting Gain is the integral value used to adjust the baseline and fine tune the rate at which the pump is allowed to move to reach the programmed setting Note For more information regarding PID controllers see Appendix B Revision 2 8 55 July 15 2015 In Inverse Mode the setting for the diastole and systole cycles are reversed Other wise the parameters are the Run Mode j M l Pulsatile Cycle Inverse Mode xi same as in Standard Mode Please select the Operation Mode you wish In Reverse Mode the systole is Please select the Operation Mode you wish operated on during the delivery Fuse Mode phase The diastole is operated on Pulsatile Cycle ReverseMode during the fill phase Systole 30 000 Revision 2 8 56 July 15 2015 Start Mode Selector allows the user has the option to set a time de lay before the pump starts running at the rate at which it has been set or can opt for the start immediately setting Manual Device 1 3 0 6300 ml min Mode Constant Rate Flow Rat
19. the size and type of the organ being studied and the nature of the experiments being performed Chamber Nut Right Angle 14 1030420 Bubble Trap 14 1030297 1031145 Fitting 10 1030460 10 1030450 Universal Plug 14 1030410 10 1030470 Luer Fitting 1032584 Elevator 14 1030753 O Ring 14 1032707 10 1030785 10 102709 Revision 2 8 26 July 15 2015 14 Chamber Lid Lung System Pressure Temperature Readings Probe Heart System B Pressure Temperature Readings Probe Revision 2 8 27 July 15 2015 Pressure Transducer Setup underside of chamber lid 3 i j Mes 3 8 Barbs use L S 18 PharMed tubing Fitting Tapered Luer 1032584 Note For 10 chamber barb size is 1 4 Use L S 17 PharMed tubing 1 Attach proper size tubing to barb on the underside of chamber Use specialized clear fitting shown below to provide a side pathway to the pressure transducer ports With Luer to barb fittings connect tubing to side pressure port at bottom of chamber lid see bottom right image A Straight Fittings w Luer Port for Pressure Transducers 1 4 1030904 3 8 1030899 1 2 1030905 2 Using another Luer to barb fitting attach to corresponding swabble port located on top of the chamber lid and drape another piece of tubing that is attached to the Pendotek pressure transducer via a female Luer to barb fitting Cut the tubing such that the pressure transducer can be taped to the
20. there is no limit Fli Direction The Flip Direction switch up allows the flow to be reversed on J every other cycle 10 repeats with flip direction switch up would result in half of the cycles 5 in the forward direction and half 5 in the reverse direction Switch down and all cy cles in the same direction Please select the Repeat Mode you wish eration RepeatMode Count allows you to set the number of en iterations Keep Pump Count Running Change by allows you to set how much to 2 change the speed of the pump for each Jj time the step is repeated Change by Max Value Max value is the maximum flow in ml min 199 300 this is limited by the tubing Keep Pump Running allows the pump to continue to run as the final flow rate is reached Revision 2 8 60 July 15 2015 Save a New Step an e Device 1 A green Plus symbol will appear below the Create step key when no steps have been created Clicking on the icon will result in a new window being opened asking to create a new step with the current C conditions Select a Unique name You cannot name a 0 000 ml min step with a preexisting name This is done to protect T Mode Constant Rate existing steps from being altered In order to reuse a name you must manually delete the cC 3 ES E _ create Step lier step name from the HART directory in TN 4 P 0 000 ml min the my documents folder Would vou li y
21. 14 PharMed tubing Peristaltic Pump 3 1 16 1 6mm ID 316 4 8mm Wm OD AS VA 16 PharMed tubing Gas in 1 8 3 1mm ID 495 1 4 6 4mm OD gt Oo 25 PharMed 4 484 tubing Gas out 3 16 4 8mm ID O2 Sensor 5 16 7 9mm CO 5 7 D pH 17 PharMed tubing 1 4 6 4mm ID 3 8 9 5mm OD e Heater with Reservoir inside 18 PharMed tubing 5 16 7 9mm ID pem Organ Chamber 7 16 11 1mm OD Note n this setup the organ is being cannulated in two locations Y fitting The tubing pathway shown through peristaltic pump 4 is optional but 3 16 Y barb the user should be aware that some of the media will drip through the Y fitting mE organ at collect at the bottom of the chamber In order to keep these 1 4 Y barb levels from getting too high this pathway and pump may need to be utilized The media is pumped back into the reservoir via the Quick LON Connect outlet ports located on the bottom of the chamber Single Channel Pump Head It is also possible to cannulate the organ in a single location in which case the tubing pathway associated with peristaltic pump 2 may e eN become unnecessary Dual Channel Pump Head Revision 2 8 30 July 15 2015 The following is an outline of potential usages of the ports on the chamber lid The system is designed to accommodate a wide variety of protocols Each user s setup will va
22. 23 2 C Logging rate allows the frequency of the data collection to be selected Logging Locati y Do ents i aes ee Mc Note collecting 4 channels of data every minute requires 1 Megabyte of oq Naat storage each day 0 05 second Flow Inp 0 5 second Pressure Inpuj v 1 second Logging Location My Documents Le 5 seconds emperature Inp 10 secondi Logging Rate 1 second I Optional Inpu 15 seconds 30 second Flow I nput allows external FlowInput Flow Rate 1 minute inputs to be associated with Flow Rat 7 each pump such as external Pressure Input ain i nalog Input flow meters emperature Input g inp Analog Input 2 Flow Rate 0 000 ml min Optional Input k EE mm Ha The selection choice will be XE Analog Input 3 oem m displayed in the Flow Rate ar Analog Input 4 Temperature 233 C ea and included in that de vice s log files Flow Rate 0 000 ml min Pressure 3 1 mm Hg Temperature 233 C Revision 2 8 48 July 15 2015 Pressure I nput allows external inputs to be associated with each pump such as the standard pressure meters or other external devices Logging Location My Documents I Logging Rate lsecond A The selection choice will be displayed in the Pressure Rate area and in Flowlnput FlowRate cluded in that device s log files Pressure Input Pressure 2 emperatureInput Pressure 1 Pri e Optional Input dean Pressure 3 Pressure 4 Analog Inpuf 1 Analog Ingat 2 Anal
23. 34 0 360 380 396 Time sec Revision 2 8 63 July 15 2015 PROTOCOL BUI LDER is a tool that is used to combine steps to form a complete protocol zz Pumps Heaters System Step Editor he Protacol Builder Ektop I Protocol Name is used to identify and Imaging SBS Iz recall protocols that have been saved TDMS Viewer kona E Protocol Description allows the operator Flow Input Flow Rate to use text in any manner that s he desires PressureInput Pressured hi to describe the protocol l i inni l Select allows the user to Step Movers al e s A choose a step to be execut low the operator td ed move steps either Frotocot Rer Eloy up or down inside a uS 7 n Number refers to the pump protocol Note the number 1 4 being select operator should be ed careful as the steps will be implement ed in time order but can be dis played out of or der J Time On will turn the pump on at a specific time and Time Off will shut it down at a specific time in the pro tocol l e pick Step SAMPLE and run Pump 1 from 0 min to 4 min NOTE The order of the step Load Protocol allows you to is dictated by the time selected The user load a currently saved protocol to can display the steps in an order that be modified and saved as anoth does not reflect when they will be execut er file name ed TIMES ARE IN MINUTES To start at 90 V seconds a step would have a time start of 1 5
24. 80800 oo File Edit Operate Tools Window Help E M S al Protocol Builder Editor On Time mins 0 PromptMessage Prompt Mode Revision 2 8 66 User Prompt allows a prompt to be displayed with out shutting the pumps off or to actually stop the pumps when the prompt is displayed Keep Running allows the prompt to appear without stopping the pumps Press ing the Keep Running This is the default Pause allows the pumps to be stopped when the prompt appears Keep run ning is the default Prompt Message provides a manner to enter text that will be displayed on the screen July 15 2015 Protocol Step Details Please select step Details Double click on the Select button Device Type Protocol Step Details cum Step Name Manual Zok Constant Rate 9 Start Immediately C Stop Never User eo nde ots You have a choice of either forcing a user prompt 2 No Repeat at a set point or selecting a device to perform an action Insert Dee men med Protocol Step Details Please select step Details Device Type Device Number Step Name Device xi Select x1 Manual I ns On Time mins Off Time mins zi AP Constant Rate 0 0 G Start Inmediately Click on Device and select the pump that the first StepMessage e Stop Never User step of this protocol is associated with 9 no Repeat Insert Protocol Step Detail
25. Flow Input Flow Rate Pressure Input Pressure 1 l emperature Input Temperature 1 A File Contents section mm rs Se allows selection of File contents oe Properties Values table Analog values graph fe 1073120141042 49 tds what part of the log N fie femen 5 to view woe ra FF Fie Viewer Settings D Values Table lists all Pure PE IE eee UU Rinder are trying to load a large amoun Loading and dis in al a can take up to logged values for the Mn pin ss wanted b reco dat ah es ridere Pon selection 2 TEES Settings sets the COE Load all values range for the values En As Load limited number of values Number of values Starting at to be displayed Hr ne EG EX Go To allows you to Fe jump to a specific it tele Cancel Help Export Table will o n save an ASCII a mum comma delimited fila Settings Loaded values 763 POR Export Table Revision 2 8 85 July 15 2015 Analog Values Graph plots the numerical data if present of the selection n ae M SS NN Ee mw a qa d IU MS File Vi er Values table Analog values graph 1 07312014 10 42 49 tdms Pump 1 FSI Legend m 5 47 13 004 PM 3 28 2013 Loaded values 0 18446744073709551615 Copy Data to Clipboard Save Graph Image i i 5 30 00 000 PM 5 40 00 000 PM 3 28 2013 3 28 2013 Settings sets the ra
26. For a small animal system the relevant sizes are as follows 1 16 male lure to barb connector 1032294 1 8 male lure to barb connector 1032295 3 16 male lure to barb connector 1032296 E20 July 15 2015 Revision 2 8 4 f more ports are needed inlet outlet adaptors can easily be built and utilized These parts are avail able within Small Animal Tubing Set kits provided by HART Inlet Outlet Adaptor for 500mL 1L amp 2L Reservoir Bottle Allows more tubing connections to be made Clear Nut 1031141 Cap 1031140 T Fitting 1032597 Green Nut 1031142 Red Nut 1031143 Note Typically green caps indicate inlets to the reservoir red caps indicate outlets 5 Use the appropriately sized female Luer to barb connectors to attach tubing to the inlet outlet adaptors The sizing Is as follows 1 16 female lure to barb connector 1032297 1 8 female lure to barb connector 1032298 3 16 female lure to barb connector 1032049 Note Caps should be used on the inlet outlet adaptors to cover any ports that are not currently participating in a flow path see image above right In this way the adaptors can be built tall enough to ensure a sufficient number of ports even if protocol changes to require more ports to be used Revision 2 8 39 July 15 2015 Chapter 4 Operating Instructions Prior to Starting Assure that all the requirements identified in Chapter 2
27. PM Device 2Parameter Changed 12 44 33 PM Device 2 9 Rate of 200 00 ml min 12 44 42 PM Start Device 3 Rate of 12 00 ml min E gt edi 12 44 44 PM Start Device 4 Rate of 12 00 ml min D 100 0 ml min 12 44 48 PM Device 3 9 Rate of 150 00 ml min Mode Constant Rate 12 44 54 PM Device 4Parameter Changed 12 44 59 PM Device 4 9 Rate of 300 00 ml min Flow Rate 0 000 ml min Flow Rate 0 000 ml min Flow Rate 0 000 ml min Flow Rate 0 000 ml min 124524 PM Stop Device 4 12 45 31 PM Stop Device 3 Pressure 1 6 mm Hg Pressure 3 4 mm Hg Pressure 4 5 mm Hg Pressure 43 mm Hg 12 45 32PM Stop Device 2 Temperature 20 9 C emperature 20 9 C Temperature 20 4 C Temperature 35 8 C System Status EE LLLCLLILGIGIIIILLA Temperatures Pressures Flow Mg Device 1 Cr Device2 Charts Device3 Charts Device4 Charts AnalogInputs User User2 User3 User4 200 V Pressure1 A t t Pressure mmHg et ri UCL 20 50 LCL 10 E Show Limits FI 20 1 1 1 l l 1 i 1 06 22 PM 1 06 32PM 1 06 40PM 1 06 48PM 1 06 56PM 1 07 04PM 1 0712PM 1 07 20PM 1 07 28PM 1 07 36PM 1 07 44 PM 1 07 52PM 1 08 00PM 1 08 08 PM_ 1 08 16 PM 1 08 22 PM Time HH MM SS 300 vi Flow1 AN ao E 200 E 150 wv amp 100 UCL 200 50 cz LCL 100 20 EMT 4 1 1 i 1 i i i i i i 1 I i Show Limits 1 06 22 PM 1 06 32PM 1 06 40PM 1 0648PM 1 06 56 PM_ 1 07 04PM 1 0
28. Rate Flow Rate 0 000 mi min 4 44 mm Hg 33 6 C 200 0 ml min Mode Constant Rate 150 0 ml min Mode Constant Rate e a a o Cat Flow Rate 0 000 ml min 4 4 mm Hg 20 5 G Flow Rate 0 000 ml min 3 4 mm Hg 21 6 C Flow Rate 0 000 ml min 1 9 mm Hg 21 7 C Pressure Pressure Temperature Pressure Pressure Temperature Temperature Temperature Temperatures Pressures Flow Rates Device 1 Charts Device 2 Charts Device 3 Charts Device 4 Charts AnaloNagyts Userl User 2 l User3 Userd No Plots 100 amp A 50 UCL 50 LCL 25 0 fg 20 1 I 1 I 1 1 I I 1 vos 5 59 45 PM 5 59 46 PM 5 59 47 PM 5 59 48 PM 5 59 49 PM 5 59 50 PM 5 59 51 PM 5 59 52PM 5 59 53 PM 5 59 54 PM_ 5 59 55 PM 5 59 56 PM 5 59 57 PM 5 59 58 PM 5 59 59 PM 6 00 00 PM Time HH MM SS No Plots EM 100 amp ft 50 UCL 50 LCL 25 w Ei E I I LU LU I I I LU I I 1 I I LU I Show Limits 5 59 45 PM 5 59 46 PM 5 59 47 PM 5 59 48 PM 5 59 49 PM 5 59 50 PM 5 59 51 PM 5 59 52 PM 5 59 53 PM 5 59 54PM 5 59 55 PM 5 59 56 PM 5 59 57 PM 5 59 58 PM 5 59 59 PM 6 00 00 PM Time HH MM SS ZomQ 33H 1W R User 1 2 3 amp 4 Charts are completely customizable ORCA provides standard displays for temperature flow rates and devices that are set The USER tabs allow an oper
29. aus tomatically adjust the electric signal to the tem perature values 23 Press Up amp Enter SERIES 16A 23 Press index to reach Set point screen 23 Press down arrow to reach Input Screen SERIES 16A 23 Press index to home screen Revision 2 8 108 July 15 2015 Setting The Desired Temperature Press the index at least once to arrive at the Set Point Screen 2 Using the Up or Down arrows adjust the value Wikke to the desired temperature fore example 37 C UJ Press enter to save this value 4 Press the Index button to return to the home screen 5 The PID will begin adjusting the temperature to the set value Revision 2 8 109 July 15 2015 Learning Mode 1 Press both the Up and Enter buttons for 1 second and when released the following screen should ap pear 2 Press the index button to display the learning screen Depending on wheth er the unit has been programmed before or it is a new unit one of the two screen Will appear If the unit has already been through a Self Train cycle It may jump directly to the next step SELF 3 Press the UP arrow until Nor or Pid changes to SELF 4 Press Enter to save setting Revision 2 8 110 July 15 2015 5 Press the index button until the follow screen appears The screen may jump to YES mode directly 6 Press the Up button until the No changes to YES SERIES 16A 7 Press enter t
30. is cut and discarded and that fresh tubing is slid onto the barb Ty wraps may also be used on barb fittings 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 Revision 2 8 40 July 15 2015 4 2 ORCA Controller Software Power Settings It is critical that the laptop power settings not be selected which will shut down the system after a period of time to save power You can select to shut down the monitor but not the actual unit Otherwise you may freeze the system in the mid dle of the run This should be the factory default setting when the ORCA was shipped To check Right click on the battery or Power Icon in the ZEE i i IP Desktop mode battery charge lower right hand portion of the screen and select Rf Del Extended Battery Life Options Power Options Dell Command Power Manager iv Adjust screen brightness H2AM 41920AM 419 28AM 419 36 Pawer Options Zoom Q Windows Mobility Center Turn system icons on or off T BB 2 23 2015 NT Bechion mic mide batten ae rge B oa Extended Battery Life Options I Del Command ommand Power Manager Power Options Adjust
31. is displayed when using the ORCA software amp 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 4 Check the following Windows auto updates is turned off otherwise it could cause the computer to restart whilst running an experiment The ORCA power cord is plugged into the ORCA controller and the wall The Communications cable is plugged into computer and into the ORCA controller The ORCA controller is turned ON Your computer s static IP should be set to 10 22 51 65 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 ORCA YOU WILL THEN HAVE TO MANUALLY CHANGE THE IP ADDRESS BACK TO 10 22 51 65 the subnet mask should be 255 255 255 0 Revision 2 8 112 July 15 2015 Heater Temperature Regulation 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 introd
32. outside of the chamber at organ level July 15 2015 28 Revision 2 8 Reservoir Configuration I nstructions 4L bottle Lung System To oxy 1 4 Sensor To oxy Sensor 3 g From Sensor From Sensor N 1 4 To Aorta VW 3 a To PA w ww gt S b Trachea z Kis E 1 8 TT From oxy 1 4 From PV au From oxy Thermocouple Thermocouple Attach swabble port or Touhy Borst Underside Please note that you should not attempt to flow gt 700 mL min through a Luer lock fitting or the pressure will be high The large ani mal reservoir bottle above is the unit of choice for large flow rates Revision 2 8 29 July 15 2015 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 2 KEY
33. peristaltic pump it is recommended that you set a convention for flow direction that you follow for all pumps i e forward flow is always designated as flow from left to right This will help avoid potential confusion Revision 2 8 22 OXYGENATI NG CARTRI DGE For solid organs it is important to maintain a 5 796 CO Air ratio in order to allow the pH to be kept at optimal levels For hollow organs silicone tubing running in a CO incubator can provide enough gas exchange For solid organs when the cell mass increases a cartridge is generally re poured 4 MA A typical cartridge is the MEDICA Italy D150 or D300 cartridges that are available from either Medica itself or Harvard Bioscience Note that there _ are two connections to the Intraluminal and two to the extra lumen space TI iil M ai IIT wil i I WM I UNA Fittings are available that can be placed on 16 tubing and autoclaved as a flow path The tubing sets are then added to the sterile cartridge in a laminar flow hood WINN A separate flow circuit is set up from the medium reservoir green fitting through a peristaltic pump to the inlet of the cartridge and from the outlet of the cartridge back to the medium reservoir red fitting Note it is recommended that the connections ar
34. s protocols KEY 14 PharMed tubing 1 16 1 6mm ID ag 3 16 4 8mm OD 16 PharMed tubing 1 8 3 1mm ID 1 4 6 4mm OD 25 PharMed tubing 3 16 4 8mm ID y 5 16 7 9mm OD u L T fitting r 1 16 T barb Z ON Single Channel Pump Head Zoon Dual Channel Pump Head Peristaltic Pump 3 Yom Reservoir Peristaltic Pump 1 Gas in Xue C e 4 5 1o T Gas out O AS Heater with Reservoir inside Organ Chamber Peristaltic Pump 4 Peristaltic Pump 4 can be used for specialty pur poses but is not included in this sample set up Note In this setup the organ is cannulated in two locations The flow from the organ is siphoned off using a T fitting in order to give a post organ O reading It is then returned to the reservoir The rea son that the line branches off is to decrease the accumulation of back pressure as the fluid approaches the O sensor There is also a tubing line coming from the Quick Connect port at the bottom of the chamber This line is optional and just serves to prevent the chamber from becoming too full which is normally not an is sue in this situation Revision 2 8 34 July 15 2015 Revision 4 Perfusion pressure pass through connections Perfusion inlet ports Note f desired a mini bubble trap can be used with the 5 5 chamber Utility po
35. select the Operation Mode you wish Run Mode Pressure Oscillation Standard Mode The High Pressure target is the upper pressure that the system will reach High Press Target mmHg i En The Low Pressure target is the lower pressure that the system will reach High Rate Low Rate ml min ml min 0 000 0 000 The High Rate is the medium flow rate the system will use to obtain the high pressure target Low Press Target mmHg yeast The Low Rate Is the medium flow rate the system will use to obtain the low pressure target mer J Il Alternating the flow rates used by the system will dictate u Z 9 the timing between the high and low values It is possible that the system may not reach the high limit if the flow rate is not fast enough to reach the high pres sure PRESSURE CYCLE MODE In Pulsatile Cycle Mode the program allows for a pulsatile flow to be provided Please select the Operation Mode you wish by the perista Itic pumps ETE y Standard Mod Pulsatile Cycle aa pap AEN There are five modes of operation Set Rate Mode Standard Set Pressure Made Set Rate mode Inverse Mode Set pressure mode Reverse Made Inverse mode Reverse mode Systole To 30 000 Revision 2 8 53 July 15 2015 Standard Mode the parameter BPM allows the Please select the Operation Mode you wish Run Mode beats mimicking strokes per minute to be
36. set Pulsatile Cycle Standard Mode Y Systole allows the 96 of the cycle that is deliv ering to be set This value is typically about 3596 In Set RATE Mode Systole 96 is the same as in Standard Mode Please select the Operation Mode you wish l Run Mode BPM is the total number of cycles systole Pulsatile Cycle Set Rate Mode Diastole per minute Systole Percentage is percentage that the pump will stay at the systole period Duty Cy Cle Systole 95 Systole Rate Systole rate is the flow rate in ml min that 30 000 0 000 the pump runs at for the systole period Diastole Rate Diastole rate is the flow rate in ml min that 0 000 the pump runs at for the diastole period Revision 2 8 54 July 15 2015 em In Set Pressure Mode constant pres sure is set and the pump varies the flow Please select the Operation Mode you wish to maintain the systole and diastole Run Mode pressure at a constant setting for each Pulsatile Cycle Set Pressure Mode f of the their respective times BPM is the total number of cycles F systole Diastole per minute Systole Percentage is percentage that PID Settings the pump will stay at the systole period P Gain Duty Cycle Systole Systole Pressure 0 000 30 000 0 000 I Gain Systole Pressure is the target pressure 0 000 for the systole period of the duty cycle Diastole Pressure Max Rate Min Rate 0 000 0 000 0 000 Diastole Pressure is the target pressure
37. stage regulator is connected to the pinch valve supplied And that the adjust ment knob on the pinch valve be set so that that bubbles coming into the bleach solution are at 1 2 second intervals Any faster is a waste of gas We recommend that the exit tubing from the gas side extraluminal is placed into a 1096 bleach solution to minimize any chance of contamination Care should be taken to check the exit container to assure it does not go dry Many people will choose to place a 0 2 pm cartridge filter in line to sterilize the gas entering the cartridge Cartridges such as Pall Gelman Acro 50 vent filters or Milli pore Aervent XL capsule This is not required but does add a safety element The source of the gas is not relevant as long as the 5 796 CO air is delivered to the cartridge It is rare that O is required and if so proper safety accommodations should be taken when using a flammable gas i Typical sources of the mix gas include a custom mixture of gas 5 796 CO air similar to the source for CO incubators ll In some cases the inlet source for a CO incubator can be split and sent to the oxygenating cartridge iii A basic two gas mixer can be used allowing for air and pure CO to be supplied into the mixer when custom tanks are not available Air can also be supplied from either a house source or a simple air pump Please note that is using house CO or Air beside the possi bility of adding a sterilizing cartridge provi
38. 0 July 15 2015 The protocol selected will now be loaded The devices effected will become grayed out A large green arrow will appear in the far right side of the screen This allows you to initiate the protocol selected Click on the green arrow and the protocol will begin Device 1 Logging Folder Name Start Time Total Elapsed Time gt Total Run Time 100 0 mi min 100 0 ml min 100 0 mi min 100 0 ml min anri Thi Mode Constant Rate T Mode Constant Rate Mode Constant Rate st Mode Constant Rate e e E Cat Ca lt 7 g d Total Paused Time Yip w E Flow Rate 0 000 ml min Flow Rate 0 000 ml min Flow Rate 0 000 ml min Flow Rate 0 000 ml min Protocol Status Protocol Alberto Loaded Pressure NaN mm Hg Pressure NaN mm Hg Pressure NaN mm Hg Pressure NaN mm Hg Protocol Ready Temperature NaN C Temperature NaN C Temperature NaN C Temperature NaN C Temperatures Temperature VN Zl Temperature IA Zl Temperature Zl Temperature LA Temperature C UCL 40 LCL 35 Af 0 I 1 I 1 1 1 1 I Show Limits L 8 40 50 AM 8 41 00 AM 8 41 10 AM 8 41 20 AM 8 41 30 AM 8 41 40 AM amp 441 50 AM 8 42 00 AM 8 42 10 AM 8 42 20 AM 8 42 30 AM 8 42 40 AM 8 42 50 AM Time HH MM SS Zoom Q Once initiated the green arrow will turn into a red square You can terminate the protocol by click ing on the red square Legg bur On12720rb DA 43 DE emus IS The pause butto
39. 14 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 Revision 2 8 3 July 15 2015 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 necessary for the new challenges of your life science research We look forward to working with you to develop new tools to assist you in solving the new challenges of regenerative medicine from the lab bench to the patient There are thousands of publications in regenerative medicine to stem cell research utilizing 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 utiliz
40. 2 Empty User Variable 3 Empty User Variable 4 Empty Layout Create Edit User Variable allows the user to decide if they would like a value to be determined from two in puts One example may be the pres Tab Name Input Analog Input 1 Le Apply Plot Remove Plot Default Plots Change x axis scale with zoom slider Pause chart to pan Change y axis scale by double clicking on min max values 82 July 15 2015 Calibration tab allows the sensors to be calibrated In some cases such as temperature an offset is the only parameter that gt t inputs Penns HOG can be entered It is suggested that a thermometer certified by an agency such as NIST be used to determine the offset Input Media Temp Type Offset Offset Apply 0 Offset 2 Limit Calculated Measured Value Raw Signal Chart 21 8406 Pressure calibration is typically performed in two stages where the zero is set and then a known pressure is applied and it is set as a span A single zero point may also be used as an alter native Select the desired pressure channel You can have up to four channels i i l Calculated Measured Value is the instantaneous value for the 1551 1651 pressure reading Use the Zero button to zero the actual value reading and Cal Value along with the Calibration button to calibrate pressure readings The zero is normally set by opening the transducer to air Chart Inputs Calibration In
41. 7 12PM 1 07 20 PM_ 1 07 28PM 1 07 36PM 1 07 44PM 1 07 522PM 1 08 00 PM 1 08 08 PM 1 08 16 PM 1 08 22 PM 4 Time HH MM SS Zoom Q ZEEE Geo Q Manual v 124446 PM Start Device 2 Rate of 12 00 ml min 12 44 24 PM Device 2Details Changed A 12 44 24 PM Device 2Parameter Changed WV 12 44 33 PM Device 2 Rate of 200 00 ml min 12 44 42 PM Start Device 3 Rate of 12 00 ml min E gt a 12 44 44 PM Start Device 4 Rate of 12 00 ml min n 100 0 ml min 200 0 ml min 12 44 48 PM Device 3 Rate of 150 00 ml min Mode Constant Rate 12 44 54 PM Device 4Parameter Changed nan m 12 44 59 PM Device 4 Rate of 300 00 ml min Flow Rate 0 000 ml min Flow Rate 0 000 ml min Flow Rate 0 000 ml min 124524 PM Stop Device4 12 45 31 PM Stop Device 3 Pressure 1 7 mmHg Pressure Pressure j Pressure 124532PM Stop Device 2 gt A Temperature 21 4 C Temperature Temperature Temperatures Pressures Flow Rates Analog Input 1 n Analog Input 2 Analog Input 3 Analog Input 4 UCL 6 LCL 2 2 d 1 D 1 1 1 1 1 D D 1 i Show Limits 1 14 49 PM 1 14 51 PM 1 14 52PM 1 14 53PM 1 1454PM 1 14 55PM 1 14 56PM 1 14 57PM 1 4 58PM 114 59PM 1415 00PM 1 15 01 PM 1 15 02PM 1 15 03PM 1 15 04 PM 1 15 04 PM Time HH MM SS Zoom Q hemes iS amp Revision 2 8 80 July 15 2015 Event Log will display the current event log This is a display and cannot be chang
42. 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 Pressure Sensor Troubleshooting Arbitrary Values or NaN Sensor is not connected expected behavior If sensor is connected and an alternate sensor has been tried on that channel it could be the cable connection at the ORCA controller or inside the ORCA Controller How to test cable connection 1 Ensure the connection at the back of the ORCA is secure pushed all the way in and thumbscrews tightened 2 Check pressure sensor again for normal operation 3 If the pressure sensor is still not functioning properly follow the white cable from the problem pressure channel sensor to the back of the ORCA Gently push bend twist that part of the cable near the point where all the channels enter the connector If this results in intermittent valid values there is an issue with the Pressure Sensor Cable Contact HART for replacement 4 f step 3
43. N mn EN EN _ mn mr l E mn mn NEN E NEN EN mn E mum u EN L EN NEN EN NEN Chamber level tubing Quick Disconnect Fittings w 3 Touhy Borst Connector a Fitting Luer Male Barb 25 each 8 Fitting Luer Female Barb Revision 2 8 OL ile 15 cm 3l 15 cm 4 19 cm 3l 3 cm BEEN En Teig BED TL IL 1030345 4 10 cm 1030346 3 4 30 cm 1030347 3 3 Male 1031025 3g Female 1031024 Male 1030986 1030987 4 1031059 1032294 1032295 1032996 1032300 1032297 1032298 1032299 1032293 93 July 15 2015 5 2 Replacement Parts cont Ting Bat KN i I l1 NPT Barb Pme am m l NPT 14 Barb 1030401 l NPT s Barb 1030700 1 NPT 74 Barb 1032701 Ja NPT 4 Barb 1032702 37 NPT Barb 1030502 Ja NPT 3 Barb 1031131 NPT 4 Barb 1031128 Right Angle NPT Bar 17s NPT Barb 1031132 V NPT Ys Barb 1031133 l4 NPT 2 8 Barb 1032706 37 NPT 2 8 Barb 1031127 sa la NPT l s Pipe 1031134 dis NPT Js Pipe 1032703 3 s NPT s Pipe 1032705 Revision 2 8 94 July 15 2015 5 2 Replacement Parts cont Mn CREDUNT Male 25 1031140 s Nut Clear Cap Ul Female 1032598 1031142 Green Nut 1033143 Ul 1031141 5 1032597 T Fitting Luer Slip Slip Reducers 1032068 1032050
44. Pressure mode constant pressure is set by the user and the pump varies the flow to main tain the pressure setting Pressure is entered to indicate the target pressure de Please select the Operation Mode you wish sired Run Mode Constant Pressure _ Standard Mode Max rate and min rate determine the maximum Daran reiten minimum flow rates at which the pump is allowed to oper d vy Sem ettings ate Max Rate Min Rate P Gain ml min ml min 0 000 0 000 0 000 I Gain PI D Setting P Gain is the proportional gain which dic 20m tates how fast the pump speed is allowed to change in or der to reach the set point Cancel Typical starting value is P 1 PID Setting Gain is the integral value used to adjust the baseline and fine tune the rate at which the pump is allowed to move to reach the programmed setting Typical starting value is 2 0 1 Note For more information regarding PID controllers see Appendix A Users can also choose between Standard Mode and the Average setting Standard Mode uses the raw data of each reading A reading is taken about every 25 nanoseconds If the signal is very noisy the Average setting is a better option which uses a rolling average over the last 1 second readings for PID controller feedback Revision 2 8 52 July 15 2015 PRESSURE OSCILLATION MODE In Pressure Oscillation Mode the system alternates be tween the high pressure target and the low pressure target Please
45. Start Device 1 Rate of 100 00 ml min Flow Rate 0 000 ml min Flow Rate 0 000 mi min Pressure 1 4 mmHg Pressure R 215 C Temperature 3 3 mm Hg 21 3 C Flow Rate 0 000 ml min Flow Rate 0 000 ml min 12 10 05 PM 5top Devicel 12 10 07 PM Start Device 1 9 Rate of 100 00 ml min Pressure 99 1 mm Hg Pressure 4 0 mm Hg 12 10 11 PM Stop Devicel Temperature 20 9 C Temperature 21 4 C System Status ELA Device1 Charts Device2 Charts Device3 Charts Device 4 Charts Analoginputs Userl User2 User3 User4 Pressure 1 Pressure 2 Pressure 3 EE Pressure 4 Pressure mmHg UCL 100 LCL 0O ae 20 i 1 1 i 1 I 1 1 1 i 1 i i Show Limits 12 26 09 PM 12 26 11 PM 12 26 12 PM 12 26 13 PM 12 26 14 PM 12 26 15PM 12 26 16PM 12 26 17 PM 12 26 18PM 12 26 19 PM 12 26 20 PM 12 26 21 PM 12 26 22PM 12 26 23PM 12 26 24 PM 12 26 24 PM l ex Swe Del Ram Been ien DE a a gradual shift or sine like wave Time HH MM SS Zoom Q Pressure Tab can be used to display any of the 4 pres Sure sensor readings by clicking on the check box next to tical they may overlap and be difficult to tell apart see above UCL upper control limit and LCL lower control limit 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 i
46. Zero Span Apply Enter Set Known Known Value Set Zero Value to 100 Span Sensor Calculated hlearured Value Raw Signal Chart 104545 11 5 July 15 2015 10 Press both the UP and ENTER buttons simultaneously and hold for 1 second and this screen will appear Press the up or down arrow to get to 10t P versus 1t P 11 Press the index button approx 17 times to arrive at the POSr screen 12 Press the up arrow button to change the InP Input Point to SPt Set Point 13 Press the enter button to save this setting Revision 2 8 106 July 15 2015 14 Press the down arrow until the top display reads 0 15 Press enter to save Chart Inputs Calibration 16 On the Calibration Tab press Set Zero Input Analog Input 3 Type Zere Span Calculated Measured Value 1 Press enter to save Raw Signal Chart 104545 1L5 H T in Amplitude 18 On the PID press the index to return to the set screen 19 Use the arrow up key to obtain the correct set temperature Typically 37 degrees this example shows 25 degrees 20 Press the enter button to save the temperature value LOVE CONTROLS MADE IN LI S A Revision 2 8 107 July 15 2015 Chart Inputs Calibration 21 Enter the desired temperature into the enter known value window typically 37 is C Input Analog Input 3 Type Zero Span Calculated Measured Value 0 0000 22 Press Set Span and the controller will
47. ata for example a spike gradual shift or sine like wave The Y Axis display can be changed by clicking on the upper or lower number which will turn gray and then the user can type in the desired value Changing the display value does not alter the data The Zoom selection allows a certain section of the chart to be zoomed into The Pan selection tool allows viewing of the graph First you must pause the display by pressing the chart icon on the lower left side of the chart The PAN bar will appear and allow you to pan backwards UCL 1500 LCL 100 AA 320 z Show Limits 12 4 2 VEL amp n 5 LEL 35 4 A a i i i Shea Limits 1 31 56 PM LIL BM 12240 PA 1 22 12 PM r mi fi a a t DA IS ww LG Pe LAT Pee LESS LDAP LEM eee Leer EERE Leer KEMA ORO ERN tme Hea C Revision 2 8 76 July 15 2015 GEE g ai Apps Pumps Heaters System Notification Support e Device1 i5 E eo Test7 v gt A y n d 100 0 ml min c 12 00 ml min 12 00 ml min Mode Constant Rate c Mode Constant Rate v Event Log e 120712PM DeviceiStep Changed 12 07 15 PM Start Device 1 Rate of 100 00 ml min E 12 07 40 PM Stop Device 1 12 07 58 PM Start Device 9 Rate of 100 00 ml min o 12 08 30PM Stop Device 1 O 12 09 05 PM Device 1Repeat Mode Changed 12 09 08 PM Device Parameter Changed Mode Constant Rate Cat Mode Constant Rate 12 09 10 PM Device 1Stop Mode Changed 12 09 29 PM
48. ation of that research in patients Revision 2 8 4 July 15 2015 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 Method Development software logs all modifications and operator entries along with the result that was measured with the system sensors allowing a complete review of an experiment and direct translation into a research method A method storage system makes it easy to reproduce methods and conditions across multiple experiments The image capture software allows for images to be captured in real time from cameras monitoring visible light IR UV and fluorescence other systems such as ultrasound ECHO are supported Revision 2 8 5 July 15 2015 Chapter 2 Getting Started 2 1 Warning and Caution Statements The use of a WARNI NG 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 CAUTI ON statement in this User Manual alerts you to where special care is nece
49. ator to select whatever they would prefer to view Customization begins by pressing the customization tool key on the upper right hand side of the charts Revision 2 8 81 July 15 2015 Customizing the Charts Chart Inputs Calibration Layout Single Chart Tab Mame l emperatures Chart Top Chart Plot Temperature 1 M Input Temperature 1 v Apply Plot Remove Plot Default Plots Change x xis scale with zoom slider Pause chart to pan Calibration Layout Single Chart v Tab Narr Temperatures Cha Top Chart oW Temperature 1 Temperature 1 Temperature 2 Temperature 3 Temperature 4 Add Plot Apply Plot Default Plots Change x axis scale with zoom slider Pause chart to pan Change y axis scale by double clicking on min max values Inputs Calibration Edit Input Name Input Media Temp Input Name Media Temp Default Names Create Edit User Variable Apply User Variable User Variable 1 Empty User Variable Name User Variable 1 Input 1 Input 2 Media Temp Media Temp Delete m is scale by double clicking on min max values Iv Deletean sure variance before and after an organ Once a variable is named it can be displayed on the user charts Revision 2 8 Chart tab will be displayed for all charts The actual display will change depending on which chart you are viewing when selecting the customiza
50. avable 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 f 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 Some protocols may not require the use of bubble traps Note n order to install the right angle fittings the bottom fitting must first be disassembled in order to fit through the Lid hole Swabable ports 1031081 ONLY AUTOCLAVE ONCE to maintain integrity Sterile Male Female Dual caps 1032581 Available sterile as alternatives to the swabble ports Bubble Trap Head 14 chamber 1030297 10 chamber 1030450 Right Angle Fitting 14 chamber 1030420 10 chamber 1030385 Revision 2 8 16 July 15 2015 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 1032688 Blanket Heater Assembly for 10 amp 14 chambers 14 c
51. cally these systems are expensive and reliability can be an issue If CO and pH are the critical values an alternate is to use phenol red in the medium which Will give an indication when red that the CO is within 5 796 and the pH is at 7 4 7 5 During development or validation a normal blood gas analyzer can be used to determine the right conditions to maintain gas levels 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 extra luminal space of the oxygenator is typically supplied with 95 air 5 CO This can be done with a premixed tank of gasses or a gas mixer The OKO system allows 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 Revision 2 8 113 July 15 2015 Pressure Transducers
52. 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 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 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 and P are preferred Revision 2 8 98 July 15 2015 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 tim
53. ct Communications cable to laptop and ORCA Controller Cable color may vary Connect mouse to and webcam to USB ports on the laptop 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 back of the pump and to the corresponding location on the ORCA Controller Note Some users stack pumps on top of another to save bench space It is recommended that tubing be fed into the pumps prior to stacking However when stacking the closing lever is not operable A recommended configuration is to place the pumps under the biological safety cabinet BSC and not stack them Revision 2 8 13 July 15 2015 5 Identify any extra pump heads and mounting screws that have been ordered These may not be im mediately needed for the system but should be kept in a safe place Pump heads 1031108 Single Channel 1031107 Double Channel Pump head mounting screws mounting hardware for either 2 1031067 3 1031068 or 4 pump heads 10311069 6 Connect heater power cord to the controller into heater slot 1 A
54. dd 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 For smaller bottles use a lab to set the height of the cap to just above the lip of the heater see step 17 An extension cable 1030961 is required 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 1030961 Mini T Adaptor 1031079 12 Probe 1 8 OD used in previous revisions 1031078 6 Probe 1 8 OD used in previous revisions J 1030960 12 Probe 1 16 OD Mini T 1030959 6 Probe 1 16 OD Mini T 1031080 Intra organ probe Mini T An extension cord may be needed Check the probe head connector to determine if you need a large oval extension cable or the small slot extension cable 1030961 Revision 2 8 14 July 15 2015 8 Loosen the compression fitting then add temperature probe to reservoir Be sure not to have
55. did not yield any valid readings the issue could be with the ORCA However as long as the pressure channel was previously working it is still most likely the Pressure Sensor Cable If you have received a new Pressure Sensor Cable and the problem still persists contact HART for service of the ORCA system Sensor has constant value will not change The sensor may be damaged or was incorrectly calibrated The same pressure or no pressure may have been applied when setting the zero and or span value First try recalibrating If problem persists try a different pressure sensor taken from another channel If the alternate pressure sensor works meaning the pressure channel is still functioning properly then contact HART for a replacement Pressure Sensor Revision 2 8 114 July 15 2015 Peristaltic 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 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 pu
56. e 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 1096 to 9096 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 54 Process Variable LI Percent Overshoot Set Point FL dM el Settling Time Steady State Error uen ee ee Guns ee ee ee et uam 0 L L L L L L L L L i L 00 02 04 06 08 10 12 14 16 18 20 22 24 26 26 30 Time seconds The following table may be of use when tuning the PID controller Effect of increasing a parameter independently Parameter Rise Time Overshoot Settling Steady Stability Time State Error Decrease Increase Small change Decrease Degrade Decrease Increase Increase Eliminate Degrade Minor change Decrease Decrease No effect Improve in theory if Ky small Revision 2 8 99 July 15 2015 Bioreactor Pl D 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 be
57. e 0 000 ml min Pressure NaN mm Hg Temperature NaN C Start Immediately begins the operation instantly upon pressing the start pump Please select the Start Mode you wish button StartMode Start After Time Delay delays the start Start Immediately of the pump by the time selected after 1 pressing the start pump button Please select the Start Mode you wish StartMode Start After Time Delay Time 2 Second Ramp allows the user to select the speed at which the 2 pump will obtain the target flow rate There are options Please select the Start Made you wish EE ds TM in increments ranging from very slow to very fast Start Immediately J sige s None is also an option for type of ramping Very Fast Fast Medium Ramping the flow is advised when running at a high flow Slow Very Slow mE rate Without a ramp the pump may stall Revision 2 8 57 July 15 2015 Stop Mode Selector allows the user has the option to set a time delay before the pump stops running at the rate at which it has been set or can opt for the user stop setting Manual Device 1 gt 0 6300 ml min Mode Constant Rate Flow Rate 0 000 ml min Pressure NaN mm Hg Temperature NaN C B Stop Mode Please select the Stop Mode you wish Stop Mode Stop Never User Stop Stop Never user Stop pump will continue running until the operator manually stops the system Please select the Stop Mode you wish hie Never User Sip
58. e that you wish p to use See table below for a description of the available Pump 1 Calibration tubing sizes and their corresponding flow constraints When Please select Head Cc bing Size and or add on Calibration you have selected a particular tubing size the minimum and Factor maximum flow rates for the selected tubing will automatically be displayed below Head Configuration F Calibration Factor Single Head 1 Flow Rates Tubing Size Size id 7 9 mm 031 in CAUTI ON The user must know the tubing size being Min 11 4 ml min Max 1140 ml min utilized for each pump head and verify that the correct tubing size is selected The pump cannot verify this for the user Multiple tubing sizes may be used in multiple heads placed on a single drive The pump drive will only calibrate itself as if all tubing sizes are identical If the user would like to have the same drive unit using variable tubing sizes In order to know the flow rate of the channels using different tubing sizes the user would have to measure the flow and pick smaller or larger tubing to decrease or increase the flow rate NOTE If a flow rate value is chosen that is above the allowed limit for the tubing the ORCA will post an alert message and automatically adjust the flow to the highest value allowed see chart below The minimum value allowed is zero However it is not recommended when accuracy or precision is required that values below
59. e the following 1 Intraluminal inlet bottom center of the cartridge is connected to the media reservoir outlet green connector 2 Intraluminal inlet top center of the cartridge is connected to the media reservoir inlet red con nector 3 Extraluminal inlet top side port connected from the OXY pinch valve Extraluminal outlet bottom side port connected to 16 size tubing to a flask containing a bleach solution Notes a it is recommended that the gas supply is connected to the top of the side ports Thus If any condensation occurs it is blown out of the cartridge through the bottom port Otherwise liquid can build up and wet out the fibers and make the gas exchange less efficient b Having the medium flow through the bottom and going against the gas flow is the norm How ever there has not been any data explaining why this is more efficient There does not seem to be any down side to having the medium flow upward so most systems will be set up this way Maintaining the oxygenating cartridge in a vertical position can be accomplished in many ways No specific configuration seems to work best The most economical seems to be a ring clamp and a ring stand which are available from any laboratory supply house Revision 2 8 23 July 15 2015 OXYGENATI NG CARTRI DGE Notes and recommendations a d Gas pressure should be less than 10 psi 5 7 atm It is strongly recommended if a gas cylin der is used a dual
60. ection 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 80mmHg 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 process by running the pump faster and faster as time goes by this is considered integral control I 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
61. ed The actual full RT 10 8 2014 1 15 36 PM i A Regen Event Log 12 44 16 PM 12 44 24 PM 12 44 24 PM 12 44 33 PM 12 44 42 PM 12 44 44 PM 12 44 48 PM 12 44 54 PM 12 44 59 PM 12 45 24 PM 12 45 31 PM 12 45 32 PM log is located in the system log file folder Start Device 2 9 Rate of 12 00 ml min Device 2Details Changed Device 2Parameter Changed Device 2 Rate of 200 00 ml min Start Device 3 9 Rate of 12 00 ml min Start Device 4 9 Rate of 12 00 ml min Device 3 Rate of 150 00 ml min Device AParameter Changed Device 4 Rate of 300 00 ml min Stop Device 4 Stop Device 3 Stop Device 2 System Status BEEN H HART Regen vi soc Jess es iS H RT 10 8 2014 1 15 36 PM A Regen Event Log File Apps Pumps Heaters System Notification Support Manual Device 1 Device 3 Device 4 es Device 2 i ae 12 44 16 PM 12 44 24 PM 12 44 24 PM 12 44 33 PM 12 44 42 PM 12 44 44 PM 12 44 48 PM 12 44 54 PM 12 44 59 PM 12 45 24 PM 12 45 31 PM 12 45 32 PM Start Device 2 Rate of 12 00 ml min Device 2Details Changed Device 2Parameter Changed Device 2 Rate of 200 00 ml min Start Device 3 Rate of 12 00 ml min Start Device 4 Rate of 12 00 ml min Device 3 Rate of 150 00 ml min Device 4Parameter Changed Device 4 Rate of 300 00 ml min Stop Device 4 Stop Device3 Stop Device 2 system Status o 100 0 ml min Mode Constant Rate Ca 300 0 ml min Mode Constant
62. ents log 73 July 15 2015 Heater Configuration Heaters will allow you to enable each pump File Apps Pumps i Heaters System Notification Support Heater Configuration will allow individual Heater 1 Enable temperature set points to be set as well as Manual Heater2 Enable assigning which heater sensor is associated Heater3 Enable with which heater Ww aS o 8 Device 2 Heater Configuration Is pn OF ee Heater Configuration l Heater 1 Controls Heater 2 Controls Heater 3 Controls Temperature Input Temperature Input Temperature Input Temperature 1 xi Temperature 2 xi Temperature 3 xi SetPoint SetPoint SetPoint l 3 l UR l 37 i Input control range 10 0 50 0 C Cancel B ps 4 Y LAM M AM eh dua LLL d Temperature 1 Temperature 2 Temperature 3 Temperature 4 Analog Input 1 Analog Input 2 Analog Input 3 Revision 2 8 74 July 15 2015 System Logging will allow data logging to begin Choose the option of Start Stop or Pause Selecting PAUSE will continue collecting data on the same file Pressing STOP will cause a new file to be started when data logging is resumed Protocols can be loaded or cleared File Apps Pumps Manual Ev d Device 1 Configuration Location allows selection of the me dium location to store files Pumps Heaters System Notification Support Logging Li Start Protocol bk
63. eservoir Heater 110 240V 50 60 Hz 6 Amp Note Heater power outputs are identical to inputs Physical Dimensions 10 5 27 cm H x 5 12 7 cm W x 18 75 47 7 cm Controller 4X6 x725 10 16 x15 24 x 18 42cm Weight 5 3 lbs 2 4 Kg Pump Heads 4 x 3 25 x 4 75 10 16 x 8 23 24 x 12 07cm Weight 1 5 Ibs 0 68 Kg Chamber 14 Chamber 19 2 x 20 5 x 19 2 48 8 x 52 1 x 48 8 cm Top Clearance 17 3 43 9cm Volume 33L Dimensions 10 Chamber 9 5 x 16 5 x 16 5 24 1 x 41 9 x 41 9 cm Top Clearance 13 9 35 3cm Volume 11 6L 5 5 Chamber 7 x 12 5 x 7 17 8 x 31 8 x 17 8 cm Top Clearance 9 22 9cm Volume 1 8L clearance is distance required above the chamber for bubble traps fittings and tubing Peristaltic Four 4 peristaltic pumps OR three 3 peristaltic pumps AND a blood pulsatile pump Pumps Pump Heads Single and dual channel pump heads can be placed on any pump drive 1 2 3 or 4 pump heads may be used per pump drive Isolated heads for use on the bench or in an incubator Cord length is 1 5m 5 Flow Rate 2 4 mL min 1 140 mL min per pump head depending on tubing selection Stroke vol vol B 100m 100 mL nn to 100 Stroke Min Phasing Adjustable Phase 0 Phase cc MIN cs NE 5096f total cycle Ratio Analog Temperature 1 4 250 350 C thermocouple only Pressure 1 4 500 mm Hg Analog Input 10 V ORCA 4 1 4 ORCA 3 1 1 3 Up to 4 analog inputs way be read Camera Mult
64. f 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 4 8L min instantaneously or 1 4 L min average are required This selection must be made at the time of purchase If the four peristaltic pump controller is chosen up to four 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 Revision 2 8 8 July 15 2015 Peristaltic Pumps Peristaltic pump heads are available in single and dual channel versions Single Channel up to 1200mL min head Dual Channel up to 1000mL min head Up to four 4 heads may be used on each peristaltic pump to provide flexibility in flow range as well as the number of channels required for the most challenging protocol
65. hamber 110V 1031097 14 chamber 220V 1031195 10 chamber 110V 103114 10 chamber 220V 1031194 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 Revision 2 8 17 July 15 2015 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 1031048 5 5 chamber 220V 1031193 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 EN a 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 Revision 2 8 18 July 15 2015 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 and labeled For each size of tubing cut a 1 piece and glue laterally as well as a piece glued so that the internal ID of the tubing is easily displayed This is useful because otherwise
66. 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 individual sterilization protocols It is also strongly recommended that PharMed tubing be used throughout the system It has shown to be the most resilient to the intense heat and pressure of the autoclave Tygon tubing is very susceptible to warping after autoclaving 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
67. he middle icon DISK SAVE to Presse g Sia s save this protocol Once you have saved E ursi e the protocol the Disk icon will disappear Alberto rest Protoc l for the mantel Select a 6 a J Select 3 81 a Click on the Green check mark to indicate you are done editing or creating this protocol The protocol builder editor will now disappear and return you to the main screen Revision 2 8 69 July 15 2015 File Apps Pumps Heaters System Notification Support To initiate a Protocol you have saved click on the system tab and select Protocol and then Load Logging gt A Manual Device 1 Pix m 100 0 ml min Mode Constant Rate 100 0 ml min Mode Constant Rate Er Flow Rate 0 000 ml min Flow Rate 0 000 ml min Pressure 157 4 mmHg Pressure NaN mm Hg Select the Protocol you want to load Temperature NaN C Temperature NaN C 1 m Search Protocol Library 2 Organize New folder Favorites Name Date modified Type WE Desktop dat 6 12 2015 8 04AM DAT File J Downloads Alberto dat 6 12 2015 amp 40 AM DAT File Recent Places test dat 6 12 2015 7 34AM DAT File __ Libraries Documents d Music Pictures E Videos Me Computer amp OS C Removable Disk 7 File name hd Custom Pattern dat Loc E oee EE Revision 2 8 7
68. how Limits J L 125424PM 125049PM 125598PM 125527PM 125546PM 125605PM 125624PM 125643PM 125722PM 12ST2IPM 125740PM 125759PM 125808PM 125837PM 125810 PM Time HHEMMESS mal H a File Apps Pumps Heaters System Notification Support About Cx Manual 2t i ge Device 2 Exit Device 1 Revision 2 8 43 About will tell you the information on the ORCA system you are working with This in cludes software version serial number and ORCA model configuration Exit will close the software and stop pumps and all activities In version 1 you can also exit by pressing the X close button in the upper right hand corner of the home screen July 15 2015 Natific ation Support Step Editor OO Protocol Builder Imaging TOMS Viewer ii Pumps R Device 2 Step Editor allows editing of a Step A step is a combination of operational parameters saved as a sin gle entity These include start time stop time repeat number Protocol Builder Steps can be stacked together to form a protocol I maging Allows users to select from multiple imaging devices to capture images TDMS Viewer allows viewing of log files Downloading the TDMS files allows conversion to an Excel readable spreadsheet Pumps you can start or stop all pumps simultaneously Note that the conditions set will be implemented if the pumps are enabled If they are not individually ena bled no pump activ
69. iple cameras may be connected via USB ports One camera may be viewed and logged simultaneously for single photos or video Alerts may be automatically sent via email Revision 2 8 117 July 15 2015 pdate Log 01 Oct 2013 Created update log Added thermocouple ordering information 08 Oct 2013 1 3 2 Created new example Config schematic for 5 5 chamber lung system pg29 Fixed minor error in PharMed tubing specifications chart 22 Oct 2013 1 4 1 Updated part numbers and drawings Added in Blanket Heater instruc tions Updated FAQs and troubleshooting section 01 Jan 2014 Clarification amp updated images 10 Jan 2014 Added additional pressure transducer setup information 14 Apr 2014 Removal of gas monitoring system 23 Sept 2014 Correction of part numbers to new 10xxxxx system 14 Oct 2014 Updated manual to new Revision 1 screens and functions 15 Dec 2014 2 1 Expand TMS instructions for viewing data files Troubleshooting Pressure Cable Revision 1 updates Expand Parts List 22 Jun 2015 2 Notification bug documented on Page 54 Updated information on Step and Protocol Builder and Team Viewer Update information on the oxygenating cartridge 15 July 2015 Update IP address and PID instructions Revision 2 8 118 July 15 2015 Regenerated Organs for Transplant HARVARD APPARATUS REGENERATIVE TECHNOLOGY Harvard Apparatus Regenerative Technology 84 October Hill Road Suite 11 Holliston MA 01746 1371 USA ww
70. isions the subnet mask should be 255 255 255 0 Power Cable 1032307 U S 1032308 Euro 1032309 UK see parts list for other countries ORCA4 Controller 1032090 110V 1032092 220V ORCA3 1 Controller 1032091 110V 1032093 220V Temperature Probe 1030960 12 probe 1 16 diameter 1030959 6 probe 1 16 diameter 1031079 12 probe s diameter used in previous versions 1031078 6 probe 8 diameter used in previoug versions Reservoir Sizes 1031105 500mL 1031104 1000mL 1031100 2000mL 1031099 4000mL Heater 1030250 110V 1030247 220V Note 1 8 diameter temperature probes use a 1 8 compression fitting while 1 16 diameter probes use a 1 16 Touhy Borst fitting 1031059 Revision 2 8 12 July 15 2015 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 Conne
71. it is often difficult to tell the correct size tubing needed at a glance 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 1 4 3 8 1 2 and 3 4 caps available made of ethylene propylene diene monomer EPDM They are shown below EPDM Plug pkg 10 1031112 EPDM Plug pkg 10 1031113 EPDM Plug pkg 10 3101114 EPDM Plug pkg 10 1031115 17 For better heating results a reservoir jack 1032099 should be used with the 0 5L 1L and 2L res ervoir bottles in order to raise the height of the reservoir cap to just above the top of the heater see images below Revision 2 8 19 July 15 2015 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 protocol 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 f 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 orga
72. ity will occur File Apps Eus Heaters System Notification Support Start All Stop All Manual Manual Revision 2 8 44 July 15 2015 Manual Device 1 Device 2 0 000 ml min Mode Constant Rate 0 000 ml min Mode Constant Rate e wt 3 o oO Ca Flow Rate 0 000 ml min Flow Rate 0 000 ml min Pressure 498 1 mmHg Pressure 409 2 mm Hg Temperature NaN C Temperature NaN C wf 3 Enable Disable Pump This primes a pump for use or deactivates a pump when not in use m gt Start Stop Run Starts or stops the corresponding pump Flow Rate Control These up down arrows manually adjust the flow rate 1 mL min 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 2 Flow Direction x Changes the direction that the pump rotates a right arrow indicates clockwise ii Create Step Use this to create a Step See Step Creation for additional info 15 Select Step Use this to select a Step See Step Creation for additional info o Green Light Glows when pump is on e Step Mode See Step Mode for additional info Start Mode See Start Mode for additional info Stop Mode See Stop Mode for additional info Repeat Mode See Repeat Mode for additional info s Device Settings Allows setting of individual device preferences
73. lling 9 Rising Falling a Rising Falling a Rising Falling Rising Falling 9 Rising Falling Threshold Threshold Threshold Threshold Threshold Threshold Threshold Threshold Debounce sec LJ Ready e Triggered Debounce sec Up Ready Triggered Clear Debounce sec Ml Ready Triggered Clear Alerts defines operational thresholds that trigger alert screen pop ups and text messages Status Debounce is the amount of time the valve must above or below trigger valve best on raise or fallen edge Debounce sec Ready Inggered Clear Debounce sec LJ Ready Triggered Clear Debounce sec e Ready e Triggered Clear Debounce sec o Ready e Triggered Clear Threshold is the value the trigger needs above or below e e e le fe Debounce sec I Ready e Triggered Clear Configuration is reserved for future updates and has been disabled in this version F Fial HAI EE HART Rec en vi Apps Manual What can we help you with Subject Message Attached Window Capture Attached Days Event Log Revision 2 8 Pumps Heaters System DSDx LA CLELSOQLLLLdRER amp N Notification Befreiung Contact HART Contact HART will send an email message to the customer support group at HART You can attach windows capture pictures and the ev
74. lood or other body animal fluids should be treated as potentially infectious materials Personal protective equipment should be worn at all times when using the In Breath Bioreactor to protect personnel from becoming contaminated as well as to help prevent cross infection and cross contamination Bench tops equipment and other potentially contaminated surfaces should be cleaned and disinfected according to the manufacturers and or the facility s procedures Any article used to clean potentially contaminated surfaces should be disposed of as Biohazardous Waste CAUTION Failure to use the manufacturers cleaning and disinfecting procedure could result in damage to the surface or equipment Biohazardous Waste Dispose of biohazardous waste according to local Regulatory requirements Revision 2 8 977 July 15 2015 Appendix A PI D 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 on the coll
75. low in the chamber Il F I D d li y d F j j i d r I t g I r July 15 2015 100 Revision 2 8 Setup Instructions Select the appropriate power cord and insert into the power cord plug NOTE THE REVI SI ON 0 99 PID CONTROLLERS ARE 100V ONLY The unit should ONLY be plugged into a 220V 100V transformer if the line voltage is 220V REVI SI ON 1 Retransmission PI D CON TROLLERS ARE 100 240V The voltage being supplied to the PID controller will be identical to the voltage supplied to the heater Please confirm the voltage required to the heater before connecting The easiest manner to identify the Revision 1 retransmission controller versus the 0 99 read only PI D controller is that the Revision 1 controller has a BNC connector to transmit the temperature to the ORCA controller 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 x t 12 30 5cm L X lg 6 15 25cm L x d 6 15 25cm L X hg 32cm OD 16cm OD 32cm OD 32cm OD Revision 2 8 101 July 15 2015 Programming Instructions Ree E 1 amp f f a a m Kee kk 2 E Tq P m
76. min To keep the pump running at the end of the step a negative 1 1 may be entered and the pump will not shut off at the end of a protocol Message is a non significant text field to remind the user what the step is LJ Saving a Protocol is accomplished by pressing the Save icon which appears after M changes have been made Green Check mark closes the protocol builder and return the user to the main page Revision 2 8 64 July 15 2015 Protocol Builder allows you string a series of commands steps together in a logical order To build a protocol start by clicking on the APPS tabs and then on Protocol Builder HARI Regen File MEE Pumps Heaters 5 Step Editor Imaging gt TOMS Viewer y 100 0 ml min Mode Constant Rate lt Flow Rate 0 000 ml min Pressure NaN mm Hg Temperature NaN C The Protocol Builder Editor screen will come up ga o4 4 3 Protocol Loader Builder E VAS im Protocol Name Protocol Description Protocol Details PAS Select E elect z Slet Select Select Select Select Revision 2 8 0 0 28 Protocol Builder Editor 0 0 65 0 0 Presently only time based protocol steps Future versions will support variable control of protocol step July 15 2015 H Protocol Loader uy sw ej zo etn 3 an a Ot
77. mp 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 al lows 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 If there is an issue with the peristaltic pumps concerning the backfill siphon of the reservoir into the chamber Watch how the pump rollers stop to ensure that they always occlude the tubing Revision 2 8 115 July 15 2015 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 se lection 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 latm 15psi to be applied The chambers can handle negative ventilation at several psi Revision 2 8 116 July 15 2015 Specifications R
78. n allows you to pause a rg ro protocol and then a green arrow will appear allowing you to continue the protocol File Apps Pumps Heaters system Notification Support N MA baw Edom BAIUIAM BALADAM Bebop a mem BEN Rina mapia AM posi imd bakit am Device 1 1 Once the protocol is completed you must return to the System Tab select Protocol and CLEAR to return to the home screen ECIL 100 0 ml min 100 0 ml min Mode Constant Rate Mode Constant Rate y Flow Rate 0 000 ml min Pressure NaN mm Hg Pressure NaN mm Hg Temperature NaN C Temperature NaN C T Revision 2 8 71 July 15 2015 Enable Notification is a global command for enabling or disabling all email and text alerts and notifications Enable Notification B When Enable is selected the enable se Jise fice lection command line is grayed out and vice versa Pumps Heaters System Notification Support E p Device 1 Users P Status gt Alerts Configuration Prompt User for Input a P Ze i 2 lee Bloc QUE Med atl M Notifications User List Users is a listing of the users that are eligible to receive notifi cations and their e mail and the text message email address Each carrier will have a definitive protocol AT amp T is phone num ber txt att net i e 5555555555 no dots or dashes just num bers Verizon is pho nenumber vtext com Name email Address Textblessage email Address C
79. n 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 If the windows become too scratched then replacements may need to be ordered The replacement part numbers are shown below Em nm pm 5 5 1031083 10 1031084 14 1031085 5 5 1031120 Silicone includes gasket includes gasket includes gasket membrane for ultrasound Revision 2 8 July 15 2015 Pump Head Configuration amp Function Pump Head Config Typical Function s Single Channel Head Perfusion into organ at a low medium flow rate Two Single Channel Heads combined by a Perfusion into organ for smooth high flow rate Y barb 4 Empty chamber to reservoir Four Single Channel Heads combined by a Y barb Highest flow rate Flow to oxygenator Two Dual Channel Heads 4 Flow to pre organ O sensor 4 channel Flow to post organ O2 CO pH sensors Flow from a medium container Increases the maximum flow by 1 2L min with a single rd i 3 Head Single or Dual Channel channel or 1 0L min with a dual channel 4 Increases the maximum flow an additional 1 2L min with th i 4 Head Single or Dual Channel a single channel or 1 0L min with a dual channel Tubing Teflon Translucent more rigid izing ID Sizing OD Tygon Clear flexible L S 13 1031101 L S 14 1031102 1 32 0 8 mm
80. n the data for example a spike Show limits should be clicked when the limit bars are desired to be displayed The Y Axis display can be changed by clicking on the upper or lower number which will turn gray and then the user can type in the desired value Changing the display value does not alter the data The Zoom selection allows a certain section of the chart to be zoomed into The Pan selection tool allows viewing of the graph First you must pause the display by pressing the chart icon O UCL 1500 LCL 100 A Show Limits Revision 2 8 lower left side of the chart The PAN bar will appear and allow you to pan backwards Ss a h i i L2 PM 1223 PM 1 223 PM fh 71 July 15 2015 E HART Regen File Apps Pumps Heaters System Notification Support ART 10 8 2014 12 45 28 PM l Regen Event Log Device 3 Device 1 12 44 11 PM Enabled Device 3 12 44 12 PM Enabled Device 4 12 44 16 PM Start Device 2 Rate of 12 00 ml min 12 44 24 PM Device 2Details Changed 12 44 24 PM Device 2Parameter Changed 12 44 33 PM Device 2 Rate of 200 00 ml min 12 44 42 PM Start Device 3 Rate of 12 00 ml min 12 44 44 PM Start Device 4 9 Rate of 12 00 ml min 12 44 48 PM Device 3 Rate of 150 00 ml min 12 44 54 PM Device 4Parameter Changed 12 44 59 PM Device 4 Rate of 300 00 ml min 12 45 24 PM Stop Device 4 System Status EEEEEEEEEEIIGGLGGGGIMIIILLI A
81. nge for the values to be displayed Save Graphic I mage saves a bitmap image of the graph displayed Saving Files Be certain to create an experiment folder in my documents by naming the experiment see below Experiment Name Once you save after naming the experiment a folder will be available in the com puter s My Documents folder and can be copied onto a portable drive and taken to another PC for evaluation Revision 2 8 86 July 15 2015 4 4 Image Capture System Support Imaging allows cap ture of images using various imaging devic es or microscopes Pumps Heaters System Motification Step Editor Protocol Builder TDMS Viewer Device P5 Image Acquisition DN H EN Device is used to select the imaging Device 1 integrated Webcam E LED OFF ON oN2 ff device to be used Refresh will refresh and present the list of connected devices LED will control the LEDs on the mi croscope cameras supported Capture will save the image at a point in time on the desktop You can also use the F5 button on the key board to capture images Capture Viewer allow viewing of existing captures for side by side comparisons Captured Image Recalled Image Capture F5 Capture Viewer lt lt lt Recall Image Capture 2014 10 09 10 44 57jpg Open Folder Revision 2 8 87 July 15
82. nges the actual steps Imaging on this page TDMS Viewer Device 2 Loaded Step Library Manual Testl Step Library is a listing of all the steps that are available Double click on the step to open the Step Editor Delete a step by selecting hit and pressing the RED x Save Changes Button will appear after a step has been deleted or changed Click this to confirm the changes made a re saved Green Check Mark is used to exit the screen There is no step editor function available while running If you select a specific step and then go back to manual mode the settings of the step you selected will be car ried over to the manual mode Revision 2 8 62 July 15 2015 Step Viewer Selector is available after double clicking on the step Changes can now be made or new steps defined Step Name will cause a drop down selection screen to appear listing all of the existing steps See other sections of the manual for description of the step functions Step Description is an automatically generated listing of all activities in a step Step Viewer Selector Step Name p ol ORpt3Inc100 Jly2Rate1001 RunlORpt3incl Step Description e Delay for 2 second before Starting Run a Constant Rate 100 ml min Infuse io Constant Rate Direction Run for 10Seconds Repeat 3 Times Increase by 100 E Start After Time Delay Q Stop on Time Target 20 40 60 80 100 120 140 160 180 20 0 220 240 26 0 28 0 30 0 32 0
83. ning in the background Should you need to access the system you can then use this information to gain access The session will not start until a person has logged into the system so the password and session number can remain open 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 Leert Manage Shire IP GallerySystems NITDMEXCEL 14 0 0 exe http ftp ni com pub gdc epd nitdmexcel 14 0 0 exe Installation Instructions TMS Importer Download the file NITDMEXCEL 2014 0 0 exe Double click on the file nitdmexcel 14 0 0 exe Follow the installation dialogs After the installation of the TDM Excel Add In you need to start Microsoft Excel at least once using administrator privileges 4 TMS Importer Plus How to Copy Attributes Once the program is installed it can be accessed by clicking on the ADD IN tab of the int ha Excel worksheet Revision 2 8 84 July 15 2015 TDMS Viewer allows the operator to Pumps Heaters Notification i umps Heaters System Notification ow a log file on the ORCA laptop _ Step Editor Logt Protocol Builder To select an existing log file choose the Imaging files from the listing in the window that TDMS Viewer appears
84. o save the selection The PID controller will now go into a learning mode This can take over an hour depending on the temperature of the liquid that it has to start with NOTE IT IS STRONGLY RECOMMENDED THAT THE LEARNING CYCLE BE PERFORMED WI TH WATER During this process the PID controller may begin with a range above 40 C This may denature any medium such as DMEM It is recommended that the unit learn in water and then after this is completed medium be added Revision 2 8 111 July 15 2015 Appendix B Frequently Asked Questions amp Troubleshooting ORCA 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 Problem An error message
85. og Ifput 3 Flow Rate 4nalog input 4 Pressure 2 9 mm Hg Temperature 23 3 C Temperature I nput allows external inputs to be associated with each pump such as the standard temperature probes or other exter Logging Rate 1 second m nal devices Logging Location My Documents I FlowInput Flow Rate The selection choice will be displayed in the Temperature area and s included in that device s log files Pressure Input emperature Input Optional Input Logging Location Desktop A Logging Rate 1 second I Logging Location Desktop Logging Rate 1 second v Temperature FlewInput FlowRate Optional Input allows external om inputs to be associated with each Pressure Input Pressurel Row Rate ji pump such as a gas monitoring emperature Input Temperature Pressurelnput Pressurel v system or other external devices Optional Input emperatureInput Temperature 1 lt None Optional Input Analog Input 2 The selection choice will be dis Analog inpast 1 bi played directly under the selection pake Optional Input 10 382 box and included in the device s v log files AnalogInput 3 GJ Analog Input 4 Flow Rate wv vv rr Flow Rate 0 000 ml min Pressure 1 6 mm Hg Pressure 1 7 mmHg Temperature 20 5 C Temperature 20 5 C Revision 2 8 49 July 15 2015 Pump Calibration B Pump Cabri Pump 1 Calibration Please select Head Configu
86. ollow Organ Chambers r a RE Te J a The rotating double chamber is specifically designed for cell seeding and culturing both surfaces of a tubular matrix The intraluminal and extra luminal 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 Bis ry T us Large and small animal versions are available See the hollow organ bioreactor manuals for additional information Revision 2 8 10 July 15 2015 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 1031082 Revision 2 8 Pump Cables 1031072 Pump Drive with appropriate heads 1 to 4 The type and number of heads will vary with each system 4 channel controller cable Transducers 1031074 are shipped in sealed sterile packaging Manual Pressure Calibrator 1031075 11 July 15 2015 Communications cable 1032077 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 10 22 51 59 for Rev 1 and 172 16 51 59 for older rev
87. ou like to Create a New Step with your NV mracan ranfiniiatinn 498 1 mm Hg A description is listed for reference It dg NaN C cannot be changed here Ratei00lUsmStp W 7 StenDeseriptian Start Immediately Run a Constant Rate 100ml min Infuse Direction Until User Stop Cancel Select Step Once steps are saved they are available to be selected for use If a step is selected the conditions will be loaded This allows you to start with an existing set of conditions You can make changes but to save the step you will have to rename it This is done to protect the integrity of the step so that one user does not change the conditions without another fc E i he user knowing ands using the step assuming it was as 100 0 ml min they left it originally Mode Constant Rate i9 NOTE You can define a step either in Step Builder or in is T Protocol Builder If you want to have a step generally Flow Rate 0 000 ml min available create it as a discrete step in step builder and Pressure 507 4 mmHg select it in Protocol Builder If you create a step in ee gt Protocol Builder it is only available when you are using Temperature NaN C that specific Protocol Revision 2 8 61 July 15 2015 hom Pumps Heaters System Notification Support step Editor allow the operator to EE TRA LLL edit view or delete existing steps Step Editor Protocol Builder Manual You cannot cha
88. pn x neu Windows Mobility Center Z Pie ihe compete be tipo Meer Wieder User Accounts Screen brightness p 53 ce chant plan beghinesr a 8 aera Revision 2 8 41 July 15 2015 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 Geo peristaltic pumps can be utilized quem mE Documents library Je Cen O Recent Places li HART Data i e Inid Folders HART Data and HART Data Backup _ are located in my documents The user should EE Rn never need to address these folders They are 277 B uon there as emergency backups in the event that the system files are corrupted Computer LE osic Via Network li LabVIEW Data Date modified 8 22 i File Felder Running amp Terminating the Program Be sure of the following before running the software l Windows auto updates is turned off otherwise it could cause the computer to restart whilst running an experiment 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 HARTregen initiation icon from the Desktop WARNING Shutting down the program by shutting off the PC or using any other method may result in a cor
89. pter 5 Care amp Maintenance 5 2 Replacement Parts cont 14 Chamber NEN 1030297 10 Chamber NEN 1030450 5 5 Chamber i 1031200 14 Chamber 1030420 10 Chamber 1030460 Right Angle Fitting il M i li MEAE REN lt o lug Assembly niversal Chamber t 14 Chamber 10 Chamber niversal Chamber 14 Chamber lug O Ring 10 Chamber Dual Sterile Caps oe 1032581 Revision 2 8 9 July 15 2015 5 2 Replacement Parts cont Pressure 4 Channel Cable 1031073 Pressure Transducers 00 BEEN Pressure Calibration Tool BEEN aes 1031075 NEN Media Reservoir Liner o 10 1031096 4 Nut for Chamber Lids 5 1031145 Temperature Probe 12 oo Y 1030960 Temperature Probe 6 1030959 Temperature Probe Implantable 1031080 Temp Probe Connection Cable NENNT 1030961 Peristaltic Pump Mounting Screws 1031067 mel iosios ORCA Communication Cable NENNEN Bioreactor Window amp Gasket 14 chamber 10 chamber 1031084 NEN NEN NN _ NEN 00 emer 103108 Gasket ae _ mn EE m 1032007 1031083 Elevator Assembly 14 chamber 1030753 me 1030785 Power Cable USA BENE 1032307 Power Cable Euro en 1032308 Power Cable UK en 1032309 Revision 2 8 92 July 15 2015 5 2 Replacement Parts cont NOCT zZXND TON LCS te key aes Ste ny Tapered Luer Fittinc B EPDM Plugs Masterflex Tubing Teflon Rigid Tubing EN V mn NN NE
90. put Pressure 1 I Type Zero Span I Chart Inputs Calibration he PPY Enter Input Pressure 1 i Type Select Herel Fluctua Known Known Value Set PI tions see appendix Value to 100 Span Select Input and Calibration Type a 5 fo r tro u b es h O oti n g Sensor guide Zero Span Defaults Calculated Measured Value Raw Signal Chart 1 6936 Calculated Measured Value Raw Signal Chart 1 8768 LAT TU My J m M y LN Amplitude i nn Y WARNING You must have a pressure measurement device and a way to Increase the pressure If you press the calibra tion button and do not have a way to measure the actual pressure you will establish a false reading and not be calibrate the system Manual Pressure Calibrator 1031075 Revision 2 8 83 July 15 2015 Logging in Remotely The version of Team Viewer included allows a user who is present on the system to open Team Viewer and obtain a session number and password This can then be given to a remote person who can use it by opening Team Viewer on the remote computer and entering the session number and password The remote person can then take over the ORCA controller and view or make changes to the system gas The are 2 manners to access the system remotely with no one at the system keyboard A Purchase A full version of Team Viewer www teamviewer com B Open Team Viewer and obtain a session number and password Leave the Team Viewer soft ware run
91. ration Tubing Size and or add on Calibration Factor Head Configuration Tubing Size Calibration Factor Single Head L S 18 d zi Double Heads Triple Heads Flow Rates Oud Heads T ubing Size Size td 7 9 mm 0 31 in S Min 11 4 ml min Max 1140 ml min v M Calibration Factor The number set for this parameter is the inverse of the variance of the liquid delivered The calibration factor 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 For example if the volume is set for 100 ml min and 110 ml were delivered in a minute start with a calibration factor of 0 9 and adjust after measuring Revision 2 8 50 July 15 2015 Creating Steps Click on the Apps Tab to create steps protocols Steps can only be created when the pump is disengaged Notice the Step Editor is grayed out and not available when the pump is active Rex X or green check and the green arrow indicating the zm ere ta T ag er en ern en egen Stet Pumps Heaters Sy e D ENSE step Editor i hiaging Mai Protocol Builder ES _TDMS Viewer Imaging IDM5 Viewer 100 0 ml min Run User Stop E gt 100 0 ml min Mode Constant Rate Ka 2 u
92. rtant 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 Organ Chambers D ORCA Controller E Laptop with Data Acquisition Software Revision 2 8 7 July 15 2015 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 The sizes of bottles available are 500mL 1L 2L amp AL The reservoir bottles have different size ports to accommodate a wide variety of tubing and flows Note that the flow through the Luer Lok ports may cause excessive pressure above 750 mL min and larger fittings may be necessary causing the selection of the 4L bottle Pumps Pumps are selected according to the organ and protocol requirements The ORCA controller along with your pumps of choice address the needs of the decellularization and recellularization processes o
93. rts E Mini Bubble Trap for 5 5 Chamber with Luer Fittings 1031200 Perfusion pressure pass _ through connections Perfusion inlet ports Utility ports Revision 2 8 35 July 15 2015 5 5 Chamber Lid Revision 3 Fill from reservoir Right Atrium ventricular Reservoir Pressure y pressure Extra probe ports mum Temperature probe Left Atrium ventrictifar E a pressure B us T P EM Right Atrium ventricular Left Atrium ventricu lar um Wo Bubble trap empty port Rod mounting for manipulator holder Revision 2 8 36 July 15 2015 Revision 2 8 a7 July 15 2015 3 4 Reservoir Configuration Instructions 2L 1L amp 500mL bottles 1 If not done so already cut and attach appropriate lengths of tubing to the bottom of the reservoir Cap 7 j hvu Hii I i IN Il Sant BOLL WA 2 Insert thermocouple temperature probe through a Touhy Borst connector and then through the center port on the reservoir lid so that the tip of the probe is just above the bottom of the reservoir Tighten the Touhy Borst connector until it forms a tight seal around the probe 3 Attach the appropriate sized male Luer to barb connectors to a swabable port then attach to cham ber lid for direct connections see images below
94. rupted CONFIGURATION FILE and not allow the program to initiate properly the next time it is used I mproper 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 HART Data folder 2 Copy the HART Data Backup folder into my Documents naming it HART Data NOTE Calibration information will be lost as the files are overwritten Revision 2 8 42 July 15 2015 0 000 ml min Mode Constant Fate Moda Constant Rate Flow Rate 0 000 ml min Flow Rate 0 000 mimin Flow Rate 0 000 ml min Pressure NaN mm Hg Pressure NaN mm Hg Pressure NaN mmHg Temperature NaN C Temperature NaN C Temperature NaN C Event Log 1247244 PM Device Parameter Changed Start Device 1 Rate of 40 mlman 124843 PM Device 1 Q Rate of 0 53 ml min 1248 59 PM Start Device 1 C Rate of 0453 ml min 124959 Ph Step Device 1 12 5 amp 3 PM Start Device 1 O Rate of 0053 ml min l Z5632PM Step Device 125537 PM Start Device 1 Rate of 0 53 ml min Pressure 12 57 55 PM Start Device 1 Rate of 0 63 ml min Nai mm Hg 125856 PM Stop Device T Temperature NaN C System Sots MEER Temperatures Pressures Flow Rates Devicel Charts Device2 Charts Device3 Charts Deviced Charts Analoginputs Userl User2 User3 Userd Flew Flow 2 How3 Flow 4 Flove Rate il rnin ue Ne uec um EV UCL 1500 LEL 100 S
95. ry depending on the size and type of the organ being studied and the nature of the experiments being performed Revision 4 Revision 2 8 31 July 15 2015 Revision 3 URS spare Plug Port Right Atrium Pressure 3 Left Atrium Pres H E sure 1 f zE COM S Chamber or Spare Chamber Vent Temperature Probe e amp Left Ventricle d me Pulmonary Artery ny i 20 Right Ventricle Arm Articulator holder Organ elevator handle Compression lock screw Aorta Feed Universal Plug 1030470 Pulmonary artery Right Ventricle Revision 2 8 32 July 15 2015 Quick Disconnect Fittings s 1031024 Female amp 1031025 Male Quick Disconnect Fittings 1 4 1030987 Female amp 1030986 Male For information on the 4L reservoir bottle ports refer to 14 chamber tubing setup For information on the 2L 1L or 500mL reservoir bottle ports refer to 5 5 chamber setup Refer to the 14 chamber setup for pressure transducer setup information Revision 2 8 33 July 15 2015 i 5 5 Chamber Small Animal System Example Configuration This is an example diagram representing a possible configuration for the ORCA bioreactor for a small 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
96. s HARVARD H One Single channel Two Single channel Four Single channel Head Heads Heads 1031061 1031062 1031063 One Dual channel Head Two Dual channel One each Single and 1031064 Heads 1031065 Dual channel Heads 1031066 Additional Part Numbers 1031067 Mounting Screw for Two pump heads 1031068 Mounting Screw for Three pump heads 1031069 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 Revision 2 8 9 July 15 2015 Chambers Solid Organ Chambers Large Solid Organ I ntermediate Solid Organ Small Solid Organ Chamber 14 36cm Chamber 10 25cm Chamber 5 5 14cm 1030240 holds approx 33 L 1030260 holds approx 12 L 1030320 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 sampling ports are included and the built in windows allow for manual access to an organ H
97. s Delete me mom Please select step Details Device Type Device Number Step Name Select Manual EET k Click on Device and select the pump that n ume mus EAS 1 0 CP Constant Rate the first step of this protocol is associated with 2 Start Immediately C Stop Never User StepMessage gt J No Repeat Insert Delete Cancel 67 July 15 2015 Revision 2 8 Protocol Step Details Please select step Details Device Tyne Device Number Step Name _ Device Select dl 4 Manual 1 On Time mins Off Time mins 2 0 e Stop Never User e No Repeat StepMessage Insert Delete Save Cancel EF Protocol Step Details imeem Please select step Details Device Type Device Number Step Name Device Select Manual On Time mins Off Time mins Zok Constant Rate 0 0 CS Start Immediately C Stop Never User No Repeat StepMessage ct ep 1 Insert Delete You can now select a step that is in the step library Select the time ON and OFF for the step Note if you have a step for 0 1 min and the next step starts at 2 min The pumps will shut off at 1 min and turn on at 2 min regardless if you have selected never stop in the step The protocol builder takes precedence over individual steps If you place a time of 1 minus one in the OFF Time this indicates to the Protocol builder to con tinue to run the existing s
98. screen brightness Windows Mobility Center Turn system icons on or off gu ee 994 CEE En 5 CJ F3 gt Control Panel Hardware and Sound Power Options Y 4 l Search Contro Pane nn Control Panel Home Select a power plan Require a password on wakeup Power plans can help you maximize your computer s performance or conserve energy Make a plan active by selecting it or choose a plan and customize it by changing its power settings Tell me more about power Choose what the power plans S Plans shown on the battery meter Select N EVE R on the put Choose what closing the lid does C ceri Dell Change plan settings b h Create a power plan Automatically balances performance with energy consumption on capable hardware CO m D U te r to S ee p O n ot gt Choose when to turn off the is display Power saver Change plan settings th e b atte ry a n d W h e n B Charge when the computer Saves energy by reducing your computer s performance where possible sleeps Desktop mode battery charge Show additional plans Y lj Dell Extended Battery Life v C 4 sip om 7 ce Options 1 F G Cerul Pinri Hates and eund Fene phena EF CET ta ike nge seminis for he plan Dell E hoc the erp and dapi utege Hurt vou wat y pompier in unm i Din battery a Phaggedin G Den ra depu Jean 5 rare Personalization KE Taen OT thi dipli
99. sions should be made to guard against any oil coming through the lines as well as moisture A simple feed into a side arm flask will trap any condensation and oil Some people will actually use a multi stage cartridge such as those used in gas chromatography feeds This is not necessary but does add an extra lev el of security Related Parts Oxy Cartridge D300 1 1032366 Oxy Cartridge D150 1 1032367 Media nlet Outlet Fitting 4 1032772 Gas Inlet Outlet Fitting See parts section for female Luer Barb of appropriate size Clamp 1 1032736 Stand 1 1032312 Pressure Regulator 1 1032565 Stand adapter for Pressure Regulator 1 1032560 Revision 2 8 24 July 15 2015 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 KEY 14 PharMed tubing 1 16 1 6mm ID 74 3 16 4 8mm OD 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 jv 17 PharMed tubing 1 4 6 4mm ID D 3 8 9 5mm OD 18 PharMed tubing 5 16 7 9mm ID
100. ssary 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 NOT FOR HUMAN USE 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 all times when handling and using the Bioreactor Wherever blood products are used Universal Precautions must be followed 2 4 Facility Requirements Assure that the facility is able to provide a clean safe and suitable area for aseptic cell processing It is recommend that all manipulations of the unit once sterile are performed in a biological safety cabinet laminar flow hood Failure to provide a means to conduct aseptic cell processing may result in harmful contamination Revision 2 8 6 July 15 2015 2 5 Equipment Components It is impo
101. tep until a new step is called for Therefore the pumps would not shut off between steps It is recommended that you write the step name in the Step Message field This way it will be dis played in the Protocol Builder Screen am ze _ UN E You can NOW Protocol Builder Editor repeat this pro Protocol Name Protocol Description cess for as many steps as you would like by double clicking on the next Se Protocol Details lect key l I Step 2 Select x 0 0 0 Lr Select A f 0 X E aded F Te es o 0 0 Select i i 0 0 o m Select T he 0 0 0 Presently only time based protocol steps Future versions will support variable control of protocol ster Revision 2 8 68 July 15 2015 ze me EA rn mm A 2M Protocol Builder Editor Protocol Name Pratocol Descnptton Protocol Details 197 Lm mri E l T Sem ol of o Presently only time based protocol steps Future versions will support variable control of protocol ster In this Protocol The system will run 1 Step 1 on device pump 1 from zero to 1 minute 2 Step 2 on device 1 from 1 minute until it gets another step command 3 Step 3 on device 1 will begin at the 4 minute mark and continue until the 5 minute mark 4 The pumps will shut off at 5 minutes Click on t
102. 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 The Y Axis display can be changed by clicking on the upper or lower number which will turn gray and then the user can type in the desired value Changing the display value does not alter the data The Zoom selection allows a certain section of the chart to be zoomed into The Pan selection tool allows viewing of the graph First you must pause the display by pressing the chart icon on the lower left side of the chart The PAN bar will appear and allow you to pan backwards Media Temp UCL 1500 LCL 100 Af Show Limits 1 apply 121929M 12204PM 12296PM IIMAPM L2MOTM al BEEN eal Time HEEM SS 12242 PM amp Revision 2 8 78 July 15 2015 Pumps Heaters System Notification Support File Apps Manual e 100 0 ml min Mode Constant Rate 200 0 ml min cC Mode Constant Rate ia 150 0 ml min Mode Constant Rate 300 0 ml min D Mode Constant Rate Gat 12 44 16 PM 12 44 24 PM 12 44 24 PM 12 44 33 PM 12 44 42 PM 12 44 44 PM 12 44 48 PM 12 44 54 PM 12 44 59 PM Flow Rate 0 000 ml min 3 4 mm Hg 20 9 C Flow Rate 0 000 ml min 1 6 mm Hg 20 9 C Pressure Pressure Temperature Temperatures 7 Pressurei 8 3 1 UCL 20 LCL 10 4 Show Limits F ADS Pressure mmHg 2 S I j 1 06 32 PM 1 06 40 PM v Flow 1
103. 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 1031080 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 WM il J i Ml Mi N 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 The Pendotek pressure transducers are typically taped to the outside of the chamber at organ level so that the readings are not influenced by gravity See 14 chamber lid and 5 5 chamber lid sections for additional setup instructions July 15 2015 15 Revision 2 8 Note Make sure to clean transducers thoroughly before use Pressure transducers are NOT autocl
104. the limits below are selected The pump will attempt to deliver the values but the per formance cannot be guaranteed Specifications for PharMed Pump Tubing Tubing Size 13 14 16 25 17 18 Barb Size in mm 25 2 Max Pressure periodic psig bar 3 Use in Single Channel Head YES YES Yves ves ves YES Use in Dual Channel Head yes ves Yves ves NO WO Part Number 25 foot roll 7 6m 0 8 4 8 STS 1 6 4 8 7 9 Max Pressure continuous psig bar YES S ES YES S O Revision 2 8 47 July 15 2015 Press the device settings Device Settings allows preferences of individual devices to Logging Location My Documents v be set and logged Logging Rate 1 second Logging Location allows data to Legging Location My Documents FlowInput FlowRate wl fbe saved to either a local drive a Logging Rate i esktop Pressure Input Pressure2 thumb type drive or the network if pero the IT department saves the my F documents folder automatically to Optional Input None Bthe network The location cannot be changed while logging emperature Input Temperature 2 Y emperature Input Temperature 2 I Optional Input None I Pressing this button will toggle ml min nent ml min Flow Rate 0 Flow Rate 0 000 back from the device settings Pressure 2 9 mmHg Pressure 3 4 mm Hg page Temperature 23 2 C Temperature
105. tion tool is selected If a standard chart is selected then the options are limited for change As there is only one chart on some displays the top chart is selected and grayed out Calibration Inputs If the dual chart display is selected Device 1 4 then you can select either the top or the bottom chart to change Tab name allows you to select the tab you UC a Plot ottom Chart are customizing Input Pressure 1 Layout Dual Chart Tab Name Device 1 Charts Plot allows you to select the which sensor to plot You also have an option of adding a new plot from another input Apply Plot Remove Plot Default Plots Change x axis scale with zoom slider Pause chart to pan Change y axis scale by double clicking on min max values Default Plots allows you to return to the original settings Inputs tab allows the customization of the chart displays Input allows one of the existing charts to be selected Device 3 Rate of 150 00 ml min Device 4Paramet Temperature 1 Device4 Rated Temperature 2 l Stop Device 4 Temperature 3 Input name allows the Name the side eene Temperature Stop Device 2 of the chart to be customized l e Templ can be changed to medium temp Pressure 2 Pressure 3 Pressure 4 Flow 1 Py 2 Flow 3 Flow 4 Analog Input 1 Analog Input 2 Analog Input 3 Analog Input 4 User Variable 1 Empty User Variable
106. ucing 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 Additionally 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 There are several systems available that monitor CO O and pH Typi
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Solid Organ Bioreactor Manual - Harvard Apparatus Regenerative

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