Model PDMI-2 Micro Incubator Manual
Contents
1. 2 1 39 4 16 4 2 5 45 4 22 4 2 9 49 0 26 0 This temperature difference is between the chamber and ambient positive when chamber above ambient Use this column to estimate data of chamber temperature versus current for other ambient temperatures Harvard Apparatus PDMI 2 Micro Incubator User s Manual Appendix E Thermistor Calibration Table Resistance Data PDMI 2 Internal Thermistor Temperature C Resistance KQ Temperature C Resistance KQ 4 271 6 25 100 0 5 258 3 26 95 3 6 245 7 21 913 7 233 8 28 87 4 8 2225 29 83 6 9 211 9 30 80 0 10 201 7 31 76 6 11 192 2 32 73 3 12 183 1 33 70 2 13 174 5 34 67 3 14 166 3 35 64 4 15 158 6 36 61 75 16 151 3 37 59 2 17 144 3 38 56 75 18 137 7 39 54 5 19 131 4 40 52 2 20 125 5 41 50 1 21 119 8 42 48 0 22 114 5 43 46 1 23 109 4 44 44 3 24 104 5 45 42 5 Appendix F Chamber Cable Color Codes D9 r Harvard Apparatus PDMI 2 Micro Incubattor User s Manual From the first release to the market of the PDMI 2 Micro Incubators to the current design there have been various design changes A wiring color code of the different units released is provided below to aid the wiring of the PDMI 2 cable to the chosen temperature controller s power control output In the current version the PDMI 2 is provided with a connector which matches the TC 202A output connector For other options the table below may be used In any case we suggest that you contact Harvard Apparatus I2. B Attempted operation outside recommended range if control point was chamber tempera ture and its thermistor was uncovered due to failure in per fusion system feed forward failure occurs with the TC 202A it automati cally stops delivering current Fluid creep can short this to plate Either monitor perfusion system more close ly or else switch with TC 202A to Peltier plate control Wait for Peltier plate temperature to cool off Use one of the solutions in 2 above to avoid repeated lapses in current Check for ground path between chamber ground connection and metal plates Dry agar bridge and petri dish wipe surfaces with silicone wax or oil to ensure they are hydrophobic Drift in baseline when electri cally recording is most likely due to one of Ag AgCl junc tions in complete recording cir cuit Bath thermistor is not completely disconnected If problem localized to PDMI 2 s pellet PDMI 2 should be retumed to Harvard Apparatus Inc for replacement For single channel patch clamp recording bath ther mistor should be completely disconnected for lowest noise Prior calibration will allow determining of chamber temperature from plate temperature Ground loops causing noise are avoided by using only one grounding point preferably near chamber Connect patch clamp ground bath ground white pin and most likely heat exchange plate ground green pin here Source of ground loops can be
3. better performance the feedback method is preferred In this method the current supplied is automatically adjusted in mag nitude and with the TC 202A direction depending on the difference between the actual and the desired temperature The advantage of feedback is that the calibration curve cham ber temperature versus controller setting is unaffected by ambient temperature changes In the cooling direction however the lowest temperature reachable will still depend on the degree of contact with the microscope stage and the ambient temperature Manual Control The power supply should deliver constant current rather than voltage and be adjustable up to the maximum needed for the desired temperature range Use the approximate cal ibration of chamber temperature versus current for low perfusion rates Appendix D page 17 to determine this At higher perfusion rates more current will be needed for a given temperature for static solutions less current This data will also be affected by ambi ent temperature the depth of media in the chamber and the degree of contact of the PDMI 2 with a large metal surface such as a microscope stage The data of Appendix D page 17 can be used approximately at other ambient temperatures since it is expressed as temperature difference from ambient versus current For accurate calibration produce your own graph of temperature directly measured in the chamber versus current See Setup and Operation for correct p
4. for studies of distributed synaptic inputs or network activity in which drugs need to be applied to a large area Perfusion is especially useful for the application and subsequent washout of an antagonist at a precise concentration 6 Harvard Apparatus PDMI 2 Micro Incubattor User s Manual Description Figure 1 Top View of PDMI 2 Fins 1 0 Cable Chamber Optical Window Ch 1 Perfusion Lines Salt Bridge Well 8 Ch 2 Perfusion Lines Gas Inlet 9 Plate Ground Socket green Surface to Hold Magnets 10 Salt Bridge Ground Socket white Xx I 7 Cover Peltier Ring Seal N Petri Dish Lower Plate or Cover Slip Dish Figure 2 Top Heat Transfer Plate of PDMI 2 Expanded View CEO Figure 3 Top Heat Transfer Plate of PDMI 2 Assembled View Controlling the Micro Incubator aE __ lt _ lt lt lt i Harvard Apparatus PDMI 2 Mao Incubator User s Manual The PDMI 2 has built in Peltier heating cooling elements Current passed in one direction will heat the enclosed dish reversed current will cool it WARNING The maximum sus tained current is about 6 amperes This corresponds to a maximum voltage applied across the two Peltier leads of about 3 volts Two methods of control are possible manual or feed back A manual system uses a DC power supply the direction and magnitude of the cur rent through the micro incubator is adjusted by hand For
5. of Peltier Devices Peltier devices are essential for rapid temperature changes or when the desired tem perature is close to ambient temperature because they can actively correct the actu al temperature whether too high or too low instead of relying on the slow passive return of a resistive heater system These advantages are only available when a bipo lar temperature controller is used see Section III Peltier devices also offer the flexi bility to cool as well as heat in the same chamber Such a system further allows exam ination of the temperature dependence Q 9 of biological properties by changing the command temperature rapidly during an experiment Finally lowering the tempera ture eases the study of ion channels with rapid kinetics Appendix D Temperature Current Data I Harvard Apparatus PDMI 2 Mao Incubator User s Manual Conditions Ambient temperature 23 C chamber liquid perfusion 0 5 ml min gas flow 50 ml min with cover plastic Petri dish with 1 75 ml of media Nikon Diaphot microscope no cooling water for Peltiers Current Chamber Delta amperes Temp C Temp C 4 1 9 4 13 6 3 6 10 1 12 9 3 1 10 6 12 4 2 8 5 11 3 11 7 2 56 12 1 10 9 2 3 12 6 10 4 2 0 13 5 9 5 1 8 14 5 8 5 1 55 15 1 7 9 1 35 16 1 6 9 1 15 17 1 5 9 0 95 18 2 4 8 0 75 19 4 3 6 0 55 20 7 2 3 0 35 21 9 1 1 0 15 24 1 1 1 0 35 26 0 3 0 0 65 27 8 4 8 0 95 30 2 6 8 1 2 32 4 9 4 1 5 34 8 11 8 1 8 36 4 13 4
6. subtle Check earthing system and fluid level in bubble traps Simplest way to solve problem is to use stainless steel needles in bubble traps and earth them Note See Maintenance page 12 for ways to pretreat pellet to reduce offset Harvard Apparatus PDMI 2 Micro Incubaitor User s Manual Footnotes Do 7 ZwZwmmwmq rr 1 A Forsythe I D et Al A chamber for electrophysiological recording from cultured neurons allowing perfusion and temperature control Journal of Neuroscience Meth 2d 19 27 1988 This article describes a prototype of the PDMI 2 without provision for gas flow Forsythe I D et al An open perfusion micro incubator for electrophysiological recording in vitro Journal of Physiology London 410 SP 1989 This article describes the addition of gas flow to the device and the elimination of the need for water cooling Ince C et al A teflon culture dish for high magnification microscopy and meas urements in single cells Pflugers Arch JU 240 244 1985 Purves R D ed Earthing and Interference Microelectrode Methods Intracellular Recording and Iontophoresis pages 55 65 Academic Press London 1981 DeHaan R L et al Journal of General Physiology 65 207 1975 Ince C et al Micro CO Incubator for use on a microscope Journal of Immunology Meth 60 269 275 1983 This paper describes the use of gas flow to reduce vertical and horizontal tempera
7. C T2 thermistor probe for use with the old TC 102 4 Chambers Either disposable 35 mm Corning Petri dishes or the reusable cover slip dish HAI TD HAI PTD LU CSDS can be used in the PDMI 2 Place the chosen chamber in the PDMI 2 central well Note For optimal fit with good heat exchange use the Corning 35 mm dish model 25000 Perfusion DO AW a Harvard Apparatus PDMI 2 Mao Incubator User s Manual Figure 4 LU ASP Aspirator Outlet from the chamber is by direct suction The included aspirator LU ASP was orig inally developed by Dr Can Ince University of Leiden for patch clamp applications The design of this aspirator cleverly avoids fluid level variations a source of electrical noise in electrical recording experiments The aspirator has a magnetic base to grip a matching magnetic surface see 5 Figure 1 page 6 on the top of the top plastic plate of the PDMI 2 Connect up a source of suction with a liquid trap to this aspirator The level of fluid in the chamber is determined by the height of this aspirator and can be changed with its thumbscrew The oscillations inherent in any peristaltic perfusion are easily damped with bubble traps Such traps also allow the independent chamber grounding required for low noise electrical recording Fluid Perfusion For reliability at low flow rates a peristaltic pump is preferable to gravity feed This also makes it easier to maintain fluid level on changing solut
8. Model PDMI 2 Micro Incubator User s Manual PDMI 2 Micro Incubator 65 0043 Harvard Apparatus PDMI 2 Micro Incubator User s Manual Table of Contents Warranty Information cccccccsceeeseeeeeeeeeeneeeeeeeeeeees 2 SPECIICAUONS maana 3 General Information cccessseeeeeeeeeeeseeseeeeeeees 4 Description aaa 5 6 Controlling the Micro Incubator 2 7 Setup and Operation ccsseeeeeeeesssseeeeeeeeeeeees 8 9 PEHUSION ANNA AWNAAANA 10 11 Maintenance sssiasscsenicenstassannsawtnaxanasonennnrcicdanmaiannan 12 Troubleshooting aaa 13 POOUMOIGS siensa enanar AWAN kakak 14 ADPENOLCE S hinniesnsecesncctecinnnteiuavesnassahneancandeteacanace 15 19 Appendix A Included Accessories 15 Appendix B Optional Accessories 15 Appendix C Peltier Device Use 16 Appendix D Temperature Current Data 17 Appendix E Thermistor Calibration Table 18 Appendix F Chamber Color Codes 19 Warranty and Repair Information PO Harvard Apparatus PDMI 2 Micro Incubaitor User s Manual Serial Numbers All inquires concerning our product should refer to the serial number of the unit Serial numbers are located on the rear of the chassis Calibrations All electrical apparatus is calibrated at rated voltage and frequency Warranty Harvard Apparatus warranties this instrument for a period of one year from date of purcha
9. amber Suggestion Use only one tube and wind it one extra turn one and three quarters total for the same exit point This should allow perfusions of as much as 6 ml min Once the top heat plate is removed reassembly for WARNING Do not further disassemble the PDMI 2 good seat is tricky Harvard Apparatus PDMI 2 Mao Incubator User s Manual Troubleshooting 13 FO Ta Problem Cause 1 Slow perfusion rate or excessive fluid build up in bubble trap 2 Very hot fins plate too hot to touch feed back temperature control disrupted 3 Temperature Controller not supply ing current 4 Excessive 60 Hz pick up baseline drift or noise if electrically recording 5 Static discharge during perfusion Due to excessive perfusion rate Blocked heat exchange tubing Too much heat is generated at Peltier junction nominally at ambient temperature Solution First flush tubing with 70 alcohol and then perfuse distilled water for one hour If still a problem then change Teflon tubing Resume operation after device has cooled off preferably after taking actions suggested under one of the following You have not placed device in contact with adequate metallic thermal mass or are trying to cool to too low a temperature Check if microscope stage is thermally con ducting Apply heat sink compound or sili cone oil to supporting metallic surface of microscope or install water perfusion See Appendix
10. c may occasionally require replacement Return your PDMI 2 to Harvard Apparatus for Ag AgCl disc replacement see General Information page 2 Disassembly for replacement of tubing see Figure 2 page 6 1 Remove the six screws outside rim of the cover observing carefully how the perfusion tubes are fed through their entry and exit points from the outer circular bobbin of the exposed heat exchange plate 2 Remove the tubing If better heat transfer is needed use heat sink compound for white paste zinc oxide contact an electronics repair shop to obtain some or silicone oil in the bobbin s groove Wind new tubing see Specifications page 3 through the same slots from the bobbin itself Cut the tubes to the desired lengths and thread them through those slots Reseat the plastic ring that circles the outside of the exposed heat exchange plate first rubbing a little silicone oil on its top and bottom surfaces and position the plastic plate Note For easy insertion in silicon gasket inside of 90 elbow fitting cut end of Teflon tubing at 45 3 Reattach the plastic plate with the six screws Be careful in tightening the screws to avoid stripping Optional Reassembly For faster perfusion rates lengthen the tubing on the bobbin by selecting another exit point or else reduce its wall thickness see Specifications page 3 Either allows the faster moving fluid to still get enough heat transfer before reaching the ch
11. cubator body heat exchange plates is grounded to the temperature controller chassis by the metal shell connector on the PDMI 2 cable Setup and Operation OP Harvard Apparatus PDMI 2 Mao Incubator User s Manual Two independent separate from the main cable ground pathways are also available when single channel or similar demanding electrical recordings are to be performed a Salt Bridge Ground The Ag AgCl disc in a salt bridge well see 3 Figure 1 page 6 is connected to a small 1 mm diameter white color socket see 10 Figure 1 page 6 This allows grounding of the recording chamber via an inte gral agar salt bridge b Shield Ground A small 1 mm diameter green color socket see 9 Figure 1 page 6 connected to the cable shield and aluminum components of the PDMI 2 Connecting this to a local ground can sometimes reduce 50 60 Hz interference 3 Temperature Probes If the PDMI 2 is to be driven from the chamber an external thermistor appropriate for the temperature controller in use should be placed in the chamber itself A small thermistor holder to position the thermistor probe tip into the chosen chamber optional reusable cover slip dish or plastic petri dishes is supplied with the micro incubator The thermistor holder is attached to the top of the PDMI 2 upper plate by a thumbscrew Thermistor probes available from Harvard Apparatus Inc are a The BSC T3 thermistor probe for use with the TC 202A b The BS
12. educing the quality of any electrical recordings The PDMI 2 permits control of both temperature and composition of the extracellular medium Three heat exchange interfaces to the culture s chamber allow perfusion to be stopped or started at will with minimal disturbance in temperature Application of drugs or a change in the ionic composition is achieved without disturbing the set tem perature or any electrodes The PDMI 2 accepts Corning 35 mm plastic Petri dishes or an adapter holding glass cover slips Leiden Cover Slip Dish LU CSD HAI TD or HAI PTD LU CSD S multi well dish cover slip dishes available separately from Harvard Apparatus Inc The PDMI 2 uses Peltier devices to drive the heat exchange Appendix C on page 16 describes their operation and identifies their advantages in this application The design of the PDMI 2 avoids the normal need for Peltier cooling water except at the lowest control temperatures Design Considerations 1 Compatibility with most popular microscopes 2 Provides excellent mechanical and optical access for ease of cell tissue manipulation and observation 3 Allows temperature regulation both above and below room temperature with perfusant media supplied to the PDMI 2 at room temperature 4 Allows temperature to be regulated either in the chamber bath with the external thermistor probe supplied with the recommended TC 202A tempera ture controller or with the built in thermistor useful when
13. en trally located chamber temperature to 0 2 C Heat transfer to the controlled medium during perfusion occurs largely from the tem perature driven plate to closely coupled Teflon perfusion tubing The Teflon tubes are coiled inside a circling slot groove machined in the top heat exchange plate see Figure 2 page 6 The extracellular medium first passes through these tubes and is warmed or cooled to the set temperature before reaching the chamber There are two other heat transfer mechanisms The first of these is by direct conduction from the support surface and collar to the chamber the other is through convection from air or gas flowing over the same temperature controlled plate Several different modes of operation are permissible 1 Static chamber medium at a constant set temperature Continuous perfusion at a constant set temperature 2 3 Switching perfusion of extracellular medium at a set temperature 4 Discontinuous perfusion 5 Rapid temperature changes with perfusion Why perfuse the chamber The ability to change the extracellular environment by perfusion is important for two reasons first because static experiments carried out above ambient temperature may incur rapid evaporation from small experimental chambers An oil layer at the liq uid air interface will avoid this and better insulate the system thermally and second perfusion is essential for quantitative ionic and pharmacological studies or
14. ing or rotary stages 102 75 mm x 1 78 mm H 4 046 x 0 071 H c PDMI ARO Alignment Ring for Olympus IX50 70 or IMT 2 fixed stage 109 73 mm x 1 78 mm H 4 320 x 0 071 H d PDMI FPZ Fixing Platform for Zeiss Axiovert with attachable mechanical stage e PDMI FPL Fixing Platform for Leica DAS Microscope DMIL and DMIRB E with attachable mechanical stage f PDMI ARL Alignment Ring for Leica Microscope 88 mm x 1 8mm H 8 52 x 0 072 H These rings or platforms are easily attached to the bottom plate with 3 small screws supplied with each ring or platform 2 Electrical Connections The PDMI 2 main cable has a multi pin connector at its end This connector matches the TC 202A front panel I O jack This cable provides the electrical connections for a Power to the PDMI 2 Peltier heat pumps b The PDMI 2 built in temperature feedback thermistor A table of its electrical resistance versus temperature is shown in Appendix E page 20 Note that this temperature is not exactly the same as that in the media in the dish c System ground The micro incubator s ground scheme is designed to provide the best possible noise shielding for demanding electrical recordings The PDMI 2 heat exchange plates are anodized and thus not electrically in contact with the microscope stage These plates are also electrically separated from the Peltier power leads One of those Peltier power wires is grounded within the TC 202A The micro in
15. ions A four channel pump is ideal since this allows for perfusion of several different solutions Two small 90 elbow white inputs CH1 IN or CH2 IN are provided in the micro incubator In addi tion two small 90 elbow white outputs CH1 OUT or CH2 OUT are provided for tubes that can be user directed to the desired location in the chamber Changing the perfusate can be achieved by switching between the two corresponding pump channels attached via a valve to one of the PDMI 2 s perfusion inlets Complete Layout of Liquid Perfusion For one possible complete layout of liquid perfusion see Figure 5 on next page Temperature Gradients The temperature at any particular point in the recording chamber is dependent on the mode of operation and the rate of perfusion The horizontal temperature gradient Perfusion DD Harvard Apparatus PDMI 2 Micro Incubattor User s Manual _ Peristaltic Pump gt gt Reservoirs oy Figure 5 Perfusion System depends on media perfusion rate gas flow rate depth of media and the presence or absence of a cover lid For absolute determination during a particular experiment it is essential to measure the temperature directly OUT e IN PDMI 2 PDMI 2 Perfusion Rate 1 0 ml min Static with Gas Flow and Lid with Gas Flow and Lid Figure 6 Isotherm Maps Temperature Gradients Gas Perfusion This is intended primarily t
16. nc for instructions or more details PDMI 2 Wiring Codes Wire Size Final Peltier AWG 20 Red Peltier AWG 20 Black Signal Ground salt bridge AWG 26 1 mm White connector AC Ground 60 Hz noise AWG 26 1 mm Green connector Thermistor AWG 26 Green Thermistor AWG 26 White Peltier 3 Peltier Plate Thermistor 4 Peltier 6 Peltier Plate Thermistor O Ground Shield
17. o reduce the heat loss to or gain from the local atmos phere thereby reducing temperature gradients across the dish It is possible to control the gas mixture above the perfusion chamber The gas passes over the same heat exchange as the fluids Any desirable gas mixture may be introduced into the micro incubator e g 5 CO air mixture can be used with a bicarbonate buffer medium An inlet for CO has been incorporated into the PDMI 2 Maintenance Do 7 3zZzZzZzZzZmz m mqqq rr Harvard Apparatus PDMI 2 Mao Incubattor User s Manual The PDMI 2 requires only a minimum of maintenance Periodically replace the tubing 26 ANG Teflon to avoid clogging by dust particles or growth of microorganisms With daily use replace this tubing every two months The period between changes can be increased by flushing with distilled water after use This may be followed perhaps by modest heating to dry out the tubing An occasional perfusion of 70 alcohol helps The Ag AgCl disc in the chamber ground can be pretreated for minimum DC offset and noise Pre fill the plastic well see 3 in Figure 1 page 6 in the PDMI 2 with the desired salt solution and let it soak for 2 to 4 hours A faster method is to immerse a silver wire in the filled well and apply an AC voltage between it and the white lead of the PDMI 2 A two minute application 60 Hz is sufficient with no more than a few milliamps preferably at 400 Hz or higher With frequent use the Ag AgCl dis
18. olarity of the power supply connections Feedback Control The control for temperature can be from the thermistor mounted on the top heat exchange plate built in thermistor compatible only with the TC 202A temperature con troller or from a thermistor placed in the chamber itself These locations are complemen tary in their performance Choose accordingly The plate temperature will be 1 to 2 C fur ther from ambient than the chamber temperature Its advantage is that the controller doesn t go out of control if the fluid runs out or drops below the level of the chamber thermistor The TC 202A has been designed as a bipolar and monopolar unit To achieve the feedback control and obtain maximum benefits from your PDMI 2 Micro Incubator we strongly recommend that you use the TC 202A on its bipolar mode See TC 202A setup for more information This temperature controller should be used for best results TC 202A Features 1 Allows the PDMI 2 to either cool or heat the bath preparation as well as control temperatures near ambient equally well 2 Automatically switches the current direction when the sensed temperature is higher or lower than the set point temperature 3 Allows fast changes in temperature 4 Allows 0 2 C regulation 5 Controls the PDMI 2 either with the micro incubator s internal thermistor or with the TC 202A supplied thermistor 6 Uses low ripple DC current for temperature control to allow low noise electrical reco
19. per tube for water perfusion is necessary for very low temperatures to prevent feed forward condition Place tubing in circular groove 0 45 cm wide on top of air fins Available from Harvard Apparatus Bioscience Catalog 1 4 inch OD 24 inches Long Figure 7 Copper Tubing Appendix C Peltier Device Use D6 Harvard Apparatus PDMI 2 Micro Incubattor User s Manual A thermoelectric module is a solid state device consisting of a series of semiconduc tors Most commercial devices are made from p and n doped bismuth or lead telluride The passage of electric current normally generates only heat The contradictory con cept of the Peltier effect is due to Jean Peltier who in the nineteenth century discov ered that the passage of current through two dissimilar conductors results in the junc tion either heating up or cooling down Each device consists of many couples mount ed in series and connected by copper strips sometimes overlying this is a thin piece of ceramic an electrical insulator but thermal conductor In principle they work as heat pumps that is when current is passed through the junction the device pumps heat in one direction reversing the current reverses the direction of the heat flow Thus depending on the capacities of the heat source and sink a temperature gradient is built up across the device The thermoelectric device is thus sandwiched between the object of interest and a suitable thermal mass or heat sink Use
20. rding without interference 7 Automatically shuts power off to the TC 202A no current will pass when excessive temperatures are reached so that possible system damage during feed forward fail ure is avoided see Troubleshooting page 13 Setup and Operation A Harvard Apparatus PDMI 2 Micro Incubattor User s Manual Note on Orientation Any references to right and left assume the unit is viewed as in Figure 1 from above with the electrical cable to the left see Figure 1 page 6 1 Mounting on the Microscope The PDMI 2 bottom plate has a flat surface for unrestrained mounting on various inverted microscope stages Accessories are available to lock the PDMI 2 to the stages of the microscopes from several manufacturers The locking devices come in two forms a As alignment rings that assemble to the bottom of the PDMI 2 and in turn fit on opening in the microscope stage b As fixing platforms that fit a locking mechanism of Zeis or Leica attachable mechanical stages Using a stage attachment accessory has the advantage of improved mechanical stabil ity and gives the user the ability to use the microscope stage built in X Y manipulators to position the chamber in the field of view The following microscope stage locking accessories are available a PDMI ARN Alignment Ring for Nikon Diaphot old or 300 200 TMD stage 107 75 mm x 1 78 mm H 4 242 x 0 071 H b PDMI ARZ Alignment Ring for Zeiss Axiovert glid
21. se At its option Harvard Apparatus will repair or replace the unit if it is found to be defective as to workmanship or material This warranty does not extend to damage resulting from misuse neglect or abuse nor mal wear and tear or accident This warranty extends only to the original customer purchaser IN NO EVENT SHALL HARVARD APPARATUS BE LIABLE FOR INCIDENTAL OR CONSEQUENTIAL DAMAGES Some states do not allow exclusion or limitation of incidental or consequential damages so the above limitation or exclusion may not apply to you THERE ARE NO IMPLIED WARRANTIES OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR USE OR OF ANY OTHER NATURE Some states do not allow this limitation on an implied warranty so the above limitation may not apply to you If a defect arises within the one year warranty period promptly contact Harvard Apparatus Inc 84 October Hill Road Building 7 Holliston Massachusetts 01746 1371 using our toll free number 1 800 272 2775 Goods will not be accepted for return unless an RMA returned materials authorization number has been issued by our customer service department The customer is responsible for shipping charges Please allow a reasonable period of time for completion of repairs replace ment and return If the unit is replaced the replacement unit is covered only for the remainder of the original warranty period dating from the purchase of the original device This warranty gives you specific righ
22. shallow media levels are being used Maintenance of constant low fluid level during perfusion Use of 35 mm Corning plastic Petri dishes or glass cover slips Small chamber and upstream volume to allow rapid media change ON NA NM Electrical isolation of heat exchange plates from chamber electrical ground This allows reduction of electrical noise by using a single external connection between signal ground and power ground Description SO Harvard Apparatus PDMI 2 Mao Incubator User s Manual The Open Perfusion Micro Incubator PDMI 2 is an annular shaped assembly see Figure 1 page 6 surrounding the central chamber either a disposable plastic 35 mm Corning Petri dish or the reusable Cover Slip Dish 35 mm The PDMI 2 contains two metal round plate assemblies to effect the heat transfer to the chosen chamber 1 The lower plate assembly black aluminum plate has a flat bottom surface to which the appropriate alignment ring is attached to form a mating surface with the microscope This lower assembly serves also as a radiator of Peltier pumped heat via radial cooling fins see Figure 1 page 6 on its outside diam eter 2 The top plate see Figure 2 page 6 also black aluminum supports the insert ed chamber and serves as the temperature driven plate Heat transfer to or from the chosen chamber is driven by two Peltier thermoelectric devices which when powered by a suitable feedback controller can regulate a c
23. tes Ranges between 4 um between 15 to 30 C and 20 um between 30 to 40 C Teflon tubing see 7 amp 8 in Figure 1 on page 6 Corning 35 mm Petri dishes LU CSD MSC TD or MSC PTD 0 5 to 2 0 L min 0 2 C with TC 202A at 37 C with 1 ml min perfu sion 2 C with 1 5 ml media volume 1 ml min perfusion Thermistor 100 k Max 6A Up to 3 0 ml min 17 x 152 mm 0 67 x 6 0 in 0 5 kg 17 9 oz Call Harvard Apparatus Inc for options at 25 C YSI 44011 General Information Ai Harvard Apparatus PDMI 2 Mao Incubator User s Manual The PDMI 2 Open Perfusion Micro Incubator is a unique and versatile multipurpose cell tissue culture unit which in combination with its matching temperature con troller TC 202A provides excellent control of both temperature and extracellular medium during 1 Intracellular whole cell or single channel patch recording from dissociated or cultured cells 2 Study of brain slice preparations 3 Long term optical examination of living tissue Cell culture is increasingly being used for electrophysiological and optical studies Although this preparation offers the convenience of a controlled growth environment in the incubator less consideration has been given to the microscope s environment where experimental examination takes place One reason for this is the design prob lem associated with providing environmental conditioning without limiting optical or electrode access or r
24. ts and you may also have other rights which vary from state to state Repair Facilities and Parts Harvard Apparatus stocks replacement and repair parts When ordering please describe parts as completely as possible preferably using our part numbers If practi cal enclose a sample or drawing We offer a complete reconditioning service CAUTION This apparatus is not registered with the FDA and is not for clinical use on human patients A CAUTION Not for clinical use on human patients Specifications 3 Specifications Harvard Apparatus PDMI 2 Micro Incubator User s Manual Electrical Temp Range Rate of Temp Change Perfusion Rate Thermal Expansion Plastic Perfusant Tubes Chamber Options Recommended Gas Flow Temperature Stability Temperature Gradient Across Chamber Built in Temperature Sensor Peltier Device Current Rating Media Perfusion Rates Overall Dimensions H x D Weight Microscope Stage Mounting Electrical Maximum 6 amperes continuous approx 3 V DC From 10 to 14 C below ambient to 50 C with TC 202A on conducting stage of microscope but with out supplementary water cooling of heat fins For 10 to 14 C change from 22 to 32 C perfus ing at 1 ml min into 3 ml about 3 mins with bipolar controller with Peltier plate control point Slower rates longer times would be obtained with DC supply Up to 3 ml min See Optional Reassembly in Maintenance Section on page 12 for higher ra
25. ture gradients and to control pH in an optically accessible chamber Forsythe I An environmental chamber regulating temperature and super fusion of tissue cultured neurons during electrophysiological or optical studies Electrophysiology and Microinjection Methods in Neuroscience volume 4 301 320 Academic Press New York Ed Conn PM 1991 This article provides a complete description of the final version of the PDMI 2 and its operation Appendix A Included Accessories 15 BCE eCeF Harvard Apparatus PDMI 2 Mao Incubator User s Manual 1 Aspirator LU ASP see Perfusion page 10 Plastic disposable Petri dishes sleeve of 20 Corning part 25000 Lid to close open top of chamber for longer term studies or reduced horizon tal temperature gradient The three screws on the lid are for height adjust ment only Teflon perfusant tubing If Teflon is used in the PDMI 2 add short lengths of Silastic tubing to the end s going into the chamber to allow chamber insertion and removal Teflon may be more appropriate for use of some per fusants Wires with 1 mm plugs for salt bridge and plates ground connections on one end and bare wire on the other Thermistor holder attached to PDMI 2 1 spare Appendix B Optional Accessories 1 A waterproof temperature probe to measure temperature within the chamber TC 202A This Temperature Controller comes with a suitable chamber ther mistor BSC T3 Cop
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