PDES-01AM man ver2_4 2015 - NPI Electronic Instruments
Contents
1. D Electronic f Instruments for the Life Sciences ASUS e ade Te OPERATING INSTRUCTIONS AND SYSTEM DESCRIPTION OF THE PDES 01AM PNEUMATIC DRUG EJECTION MODULE WITH ANALOG TIMER FOR EPMS 07 SYSTEMS PDES 01AM ON EJECT VERSION 2 4 npi 2015 npi electronic GmbH Bauhofring 16 D 71732 Tamm Germany Phone 49 0 7141 0730230 Fax 49 0 7141 9730240 support npielectronic com http www npielectronic com PDES 01AM User Manual Table of Contents A E E ine toy 3 2 EPMS 07 Modular Plug In YM ia 4 2 1 General System Description Operation ooooonccncccccnnnoonnnnnnnnnnnnonnnnnnnnnnnnnnnnnnnananannnnnnos 4 2 E EMISOR oe ee eee ee eee ne Ree 4 20 EEMS FEO AOS coo 4 De EPMS E O LOU STING O nuana tite awsa ct ee aeatieee oe ueunea a ea ueeeaneateete 4 De Dis ENS O asi ss e a nea alk aan aes ates ou caiee eanae eines 5 ZO IP WIR OSD oarn ii varanasi 5 Lalo SYSE Ct OUM INS eneo na 6 EPMSOVEPMS O39 A AA A AAA AAA AA 6 EPM S TOT na id ios 6 Ds NA Datira E E A 6 EPMS 0 7 EPMS E 07 and EPMS H O 7era os 6 EPMS 07and EP IVS OF aa a 6 FP VES SOT dial 6 Be FeSO coito pb nati abi rot iainesi 6 A AR 7 dale OVS teil DescrploN alcala aN 7 S325 FDESOIAM COMPONEN S rita att actnea lamination dee e a 7 J34 Description Ob the Front PAE ov siete erated aa weenie OS 8 4 MERU UIIAL ViVo hie ts 11 As ES Remo AV to A 11 4 2 MicroJECT micro Valve Pipette HOME nia ad 12 O sean E wah ousawla nada taeweiisunatadend eam2. 2 seals into the pipette connector in order to prevent the pipette from moving Figure 8 microJECT pipette insertion Caution Pipettes that are not rigidly fixed in the pipette connector might be pressed out of the connector by high pressure Always adhere to the appropriate safety measures version 2 4 page 12 5 E PDES 01AM User Manual Operation Connect the gas source max 5 bar to the INPUT connector at the front panel The gas source must be filtered by a 5 um filter element Important Do not connect sources of OXYGEN or COMBUSTIBLE gases The gas source MUST be dry and oil free Please use a degreaser and filter the gas source by a 5 um filter element E Connect the injection pipette to the OUT connector at the front panel or connect the microJECT pipette holder to the two microJECT connectors at the front panel 4 OUT and 5 EXTERNAL VALVE Figure 5 and connect your ejection pipette to the microJECT 1 Figure 6 If a low retain pressure analogous to the retaining current used in 1ontophoretic systems is required connect the retain pressure source to EXHAUST at the front panel Turn on pressure and adjust the pressure to the desired value using the pressure regulator Connect a timing unit to the TTL INPUT connector 9 Figure 5 at the front panel or connect a pushbutton to the REMOTE CONTROL connector 8 Figure 5 at the front panel The LED is green when no ejection occurs a
3. TIMING MON 200 ms if the internal timer is set to 200 ms Figure 9 Operation modes of the PDES 01AM version 2 4 page 14 6 E PDES 01AM User Manual Literature Bickmeyer U Heine M Manzke T amp Richter D W 2002 Differential modulation of Ih by 5 HT receptors in mouse CA1 hippocampal neurons Eur J Neurosci 16 209 218 Y Eggermann E amp Feldmeyer D 2009 Cholinergic filtering in the recurrent excitatory E E Ld E microcircuit of cortical layer 4 Proc Natl Acad Sci U S A 106 11753 11758 Garaschuk O Milos R I amp Konnerth A 2006 Targeted bulk loading of fluorescent indicators for two photon brain imaging in vivo Nat Protocols 1 380 386 Guzman J N Hernandez A Galarraga E Tapia D Laville A Vergara R Aceves J amp Bargas J 2003 Dopaminergic modulation of axon collaterals interconnecting spiny neurons of the rat striatum J Neurosci 23 8931 8940 Hahnel C 1992 Quantitative Microinjection in H Kettenmann and R Grantyn eds Practical Electrophysiological methods Wiley Liss New York Hanganu I L amp Luhmann H J 2004 Functional Nicotinic Acetylcholine Receptors on Subplate Neurons in Neonatal Rat Somatosensory Cortex Journal of Neurophysiology 92 189 198 Hofmeier G amp Lux H D 1981 The time course of intracellular free calcium and related electrical effects after injection of CaCl into neurons of the sna
4. earth Before opening the cabinet unplug the instrument Unplug the instrument when replacing the fuse or changing line voltage Replace fuse only with an appropriate specified type 3 STATIC ELECTRICITY Electronic equipment is sensitive to static discharges Some devices such as sensor inputs are equipped with very sensitive FET amplifiers which can be damaged by electrostatic charge and must therefore be handled with care Electrostatic discharge can be avoided by touching a grounded metal surface when changing or adjusting sensors Always turn power off when adding or removing modules connecting or disconnecting sensors headstages or other components from the instrument or 19 cabinet 4 TEMPERATURE DRIFT WARM UP TIME All analog electronic systems are sensitive to temperature changes Therefore all electronic instruments containing analog circuits should be used only in a warmed up condition 1 e after internal temperature has reached steady state values In most cases a warm up period of 20 30 minutes is sufficient 5 HANDLING Please protect the device from moisture heat radiation and corrosive chemicals HIGH PRESSURE PRECAUTIONS While working with PDES systems and components always adhere to the appropriate safety measures for handling high pressure pneumatic systems Before using the PDES or any external pneumatic device please read manuals and instructions carefully e Always turn off pressure source before connect
5. TROL MONITOR VALVE Figure 5 PDES 01AM front panel view the numbers are related to those in the text below version 2 4 page 8 PDES 01AM User Manual 1 Manometer Display indicating the ejection pressure range 0 4 bar 0 58 psi or 0 2 5 bar 0 36 psi 2 EXHAUST connector Connector for connecting a retain pressure source or for EXHAUST If no retain pressure is connected the system is exhausted as long as no pressure is applied to the pipette 3 IN connector Connector for connecting the filtered dry and oil free gas source The gas source must be filtered by a 5 um filter element Important Do not connect sources of OXYGEN or COMBUSTIBLE gases Note npi recommends an input pressure of bar 14 psi more than the maximum output pressure 4 OUT connector eee Connector for connecting the injection micropipette or for connecting the external Y MN valve e g microJECT The EJECT pressure 1s supplied to this connector 5 Pressure regulator Rotary knob for regulating the pressure of the ejection channel The pressure is indicated by the manometer 1 6 EXTERNAL VALVE connector Connector for the electrical connection of the external valve e g microJECT EXTERNAL VALVE 7 TIMING MONITOR 38 BNC connector providing a TTL signal that indicates the status of the regulated I injection valve HI valve open LO valve closed TIMING MONITOR ver
6. a H Furukawa T Fukuda A amp Luhmann H J 2007 GABA A receptors regulate neocortical neuronal migration in vitro and in vivo Cereb Cortex 17 138 148 version 2 4 page 16 PDES 01AM User Manual 7 Technical Data Source gas air nitrogen helium etc no aggressive combustible or explosive gases filtered by a 5 um filter element Pressure range input pressure max 5 bar 1 bar 10 Pa pressure control range 0 4 bar display range 0 4 bar or 0 2 bar display range 0 2 5 bar Input and output couplings hose fitting connect type for 4 mm inner diameter tubing Status EJECT time indicator red green LED red light valve is closed Gn EXTERN TIMER CONT operation mode green light valve is open Off no light valve is closed Mode selection four position toggle switch OFF EXTERN TIMER CONT Minimum pressure pulsed width 10 20 ms dependent on tubing 1 ms with microJECT Analog timer 0 001 0 1 s 0 01 1s 0 1 10s or 1 100 s with linear control range set by rotary switch time base selector x0 01 s x0 1 s x1 s x10 s TTL INPUT standard BNC connector TTL LO 0 2 5 V HI 2 5 12 V input impedance 10 kQ version 2 4 page 17
7. cates the operational status of the ejection channel red light valve is closed 1 e OUT connector is closed and EXHAUST connector is open green light valve is open 1 e OUT connector is open and EXHAUST connector is closed Off no light valve is closed 4 External Valves External valve s for the PDES system are either mounted on a remote valve plate or in the microJECT micro valve pipette holder 4 1 Remote Valve Plate The valve which is controlled by the PDES is located on the remote valve plate see Figure 6 Tubing and cable of the external valves are connected at the front panel of the PDES system refer to chapter 3 3 Exhaust is usually not connected Figure 6 external valves on remote valve plate 1 electrical connection black 2 EJECT pressure tubing red 3 connector for ejection pipette 4 exhaust connector optional version 2 4 page 11 PDES 01AM User Manual 4 2 microJECT micro Valve Pipette Holder The valve which is controlled by the PDES is located in the micro valve pipette holder called microJECT see Figure 7 Tubing and cable of the microJECT are connected at the front panel of the PDES system refer to chapter 3 3 Exhaust is usually not connected Figure 7 microJECT 1 pipette connector 2 connector for EJECT pressure 3 connector for EXHAUST pressure optional 4 electrical connection Important In experiments with high pressures gt 1 bar it might be necessary to insert
8. es Receptor Phosphorylation Desensitization and beta Arrestin Mediated Endocytosis Molecular Pharmacology 67 1434 1443 Qiu J Yao S Hindmarch C Antunes V Paton J amp Murphy D 2007 Transcription factor expression in the hypothalamo neurohypophyseal system of the dehydrated rat upregulation of gonadotrophin inducible transcription factor 1 mRNA is mediated by cAMP dependent protein kinase A J Neurosci 27 2196 2203 version 2 4 page 15 PDES 01AM User Manual I Stepanyan R amp Frolenkov G I 2009 Fast adaptation and Ca2 sensitivity of the mechanotransducer require myosin X Va in inner but not outer cochlear hair cells Journal of Neuroscience 29 4023 4034 _I Wolfram V amp Juusola M 2004 The Impact of Rearing Conditions and Short Term Light Exposure on Signaling Performance in Drosophila Photoreceptors Journal of Neurophysiology 92 1918 1927 Microinjection into Oocytes _I Wetzel C H Oles M Wellerdieck C Kuczkowiak M Gisselmann G amp Hatt H 1999 Specificity and sensitivity of a human olfactory receptor functionally expressed in human embryonic kidney 293 cells and Xenopus Laevis oocytes J Neurosci 19 7426 7433 Fast Drug Application with microJECT I Courjaret R Troger M amp Deitmer J W 2009 Suppression of GABA input by Al adenosine receptor activation in rat cerebellar granule cells Neuroscience 162 946 958 I Heck N Kilb W Reiprich P Kubot
9. es baa tatinseinsiewesaeoiseauech 13 O SCL AGUS e E 15 Microinjection Into DOES uti ao 16 Fast Drug Application with microJECT cccccnnnoonnnnnnnnnnnnnonononocnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnos 16 Te becia ak DA Sisa 17 version 2 4 page 2 PDES 01AM User Manual 1 Safety Regulations VERY IMPORTANT Instruments and components supplied by npi electronic are NOT intended for clinical use or medical purposes e g for diagnosis or treatment of humans or for any other life supporting system npi electronic disclaims any warranties for such purpose Equipment supplied by npi electronic must be operated only by selected trained and adequately instructed personnel For details please consult the GENERAL TERMS OF DELIVERY AND CONDITIONS OF BUSINESS of npi electronic D 71732 Tamm Germany 1 GENERAL This system is designed for use in scientific laboratories and must be operated by trained staff only General safety regulations for operating electrical devices should be followed 2 AC MAINS CONNECTION While working with the npi systems always adhere to the appropriate safety measures for handling electronic devices Before using any device please read manuals and instructions carefully The device is to be operated only at 115 230 Volt 60 50 Hz AC Please check for appropriate line voltage before connecting any system to mains Always use a three wire line cord and a mains power plug with a protection contact connected to ground protective
10. g In addition to the standard power supply of the EPMS 07 the EPMS H 07 has a built in high voltage power supply This is necessary for all MVCS MVCC modules the HVA 100 HV TR1I50 and HVC 03M modules The output voltage depends on the modules in use 2 4 EPMS E 07 Housing The following items are shipped with the EPMS E 07 housing EPMS E 07 cabinet External Power supply PWR 03D Power cord PWR 03D to EPMS E 07 Mains chord Fuse 1 6 A 0 8 A slow inserted Front covers SNNNNN N The EPMS E 07 housing is designed for low noise operation especially for extracellular and multi channel amplifiers with plugged in filters It operates with an external power supply to minimize distortions of the signals caused by the power supply version 2 4 page 4 PDES 01AM User Manual 2 5 EPMS 03 The following items are shipped with the EPMS 07 housing Y EPMS 07 cabinet with built in power supply Y Mains cord Y Fuse 034 A 0 2 A slow inserted Y Front covers DPA 2FS npi OFFSET Tit Figure 2 Left front view of EPMS 03 housing Right rear panel detail of EPMS 03 and EPMS 07 housing In order to avoid induction of electromagnetic noise the power supply unit the power switch and the fuse are located at the rear of the housing see Figure 2 right 2 6 PWR 03D The external power supply PWR 03D is capable of driving up to 3 EPMS E housings Each housing is connected by a 6 pole cable from one
11. il Helix pomatia Pfl gers Arch 391 242 251 Kolbaev S amp Draguhn A 2008 Glutamine induced membrane currents in cultured rat hippocampal neurons Eur J Neurosci 28 535 545 Lalley P M 1999 Microiontophoresis and Pressure Ejection in U Windhorst and H Johansson eds Modern Techniques in Neuroscience Research Springer Berlin Heidelberg New York Langer J amp Rose C R 2009 Synaptically induced sodium signals in hippocampal astrocytes in situ J Physiol 587 5859 5877 Lin B J Chen T W amp Schild D 2007 Cell type specific relationships between spiking and Ca2 i in neurons of the Xenopus tadpole olfactory bulb Journal of Physiology 582 163 175 Marandi N Konnerth A amp Garaschuk O 2002 Two photon chloride imaging in neurons of brain slices Pflugers Arch 445 357 365 Misgeld U 1992 Droplet application of Agonists in H Kettenmann and R Grantyn eds Practical Electrophysiological methods Wiley Liss New York Partridge L D Zeilhofer H U amp Swandulla D 1998 Combined Whole Cell and Single Channel Current Measurement with Quantitative Ca Injection or Fura 2 Measurement of Ca In P M Conn Ed Ion Channels Part B Methods in Enzymology 293 Academic Press San Diego Ponimaskin E Dumuis A Gaven F Barthet G Heine M Glebov K Richter D W amp Oppermann M 2005 Palmitoylation of the 5 Hydroxytryptamine4a Receptor Regulat
12. ing or disconnecting the PDES system e Any component used tubes fittings etc must be specified for the maximum possible pressure e Do not use any aggressive combustible or explosive gases Only gases such as air nitrogen carbon dioxide helium etc are allowed e Use only dry oil free filtered gases max 5 um filter is allowed e Maximum input pressure must be below 5 bar 72 psi version 2 4 page 3 PDES 01AM User Manual 2 EPMS 07 Modular Plug In System 2 1 General System Description Operation The npi EPMS 07 is a modular system for processing of bioelectrical signals in electrophysiology The system is housed in a 19 rackmount cabinet 3U has room for up to 7 plug in units The plug in units are connected to power by a bus at the rear panel The plug in units must be kept in position by four screws M 2 5 x 10 The screws are important not only for mechanical stability but also for proper electrical connection to the system housing Free area must be protected with covers 2 2 EPMS 07 Housing The following items are shipped with the EPMS 07 housing Y EPMS 07 cabinet with built in power supply Y Mains cord Y Fuse 2 A 1 A slow inserted Y Front covers Figure 1 Left front view of empty EPMS 07 housing In order to avoid induction of electromagnetic noise the power supply unit the power switch and the fuse are located at the rear of the housing see Figure 2 right 2 3 EPMS H 07 Housin
13. nd and mains ground connected We recommend that you try both arrangements to determine the best configuration EPMS E 07 POWER The 19 cabinet is connected to the CHASSIS connector at the rear panel It can be connected to the SYSTEM GROUND SIGNAL GROUND on the rear panel of the instrument see Figure 4 The chassis can be linked to PROTECTIVE EARTH by connecting it to the PWR 03D with the supplied 6 pole cable and by interconnecting the GROUND and PROTECTIVE EARTH connectors on the rear panel of the PWR 03D see Figure 3 Best performance is generally achieved without connection of the chassis to protective earth e a a SIGNAL Grounp CHASSIS Important Always adhere to the appropriate safety measures Figure 4 Rear panel connectors of the EPMS E 07 2 8 Technical Data EPMS 07 EPMS E 07 and EPMS H 07 19 rackmount cabinet for up to 7 plug in units Dimensions 3U high 1U 1 3 4 44 45 mm 254 mm deep EPMS 07 and EPMS H 07 Power supply 115 230 V AC 60 50 Hz fuse 2 A 1 A slow 45 60 W EPMS E 07 External power supply PWR 03D 115 230 V AC 60 50 Hz fuse 1 6 0 8 A slow Dimensions of external power supply W x Dx H 225 mm x 210 mm x 85 mm EPMS 03 Power supply 115 230 Volts AC 60 50 Hz fuse 0 4 A 0 2 A slow Maximum current supply 500 mA Dimensions 3U high 1U 1 3 4 44 45 mm 254 mm deep 265 mm wide version 2 4 page 6 PDES 01AM User Manual 3 PDES System 3 1 System Descri
14. nd red when the pressure is applied to the pipette Select the operation mode using the toggle switch 11 Figure 5 at the front panel e Inthe CONT position positive pressure is applied to the pipette continuously e Inthe EXTERN position the duration of the application is determined directly by the duration of the TTL signal or the time the pushbutton is pressed e Inthe TIMER position the duration is preset by the timer control 12 Figure 5 and time base 11 Figure 5 of the PDES Application is started with a TTL signal connected to TTL INPUT 9 Figure 5 or by pressing the pushbutton connected to 8 Figure 5 In position OFF of the timer control for pressure ejection is disabled e In the OFF position the PDES module is turned OFF the valve is closed LED 13 Figure 5 is also off The source pressure value is still shown at the manometer 1 Figure 5 but opening of the valve is disabled in OFF position Hint Ejection time can be monitored by recording the signal at 7 Figure 5 version 2 4 page 13 PDES 01AM User Manual CONTINUOUS mode TIMER CONT CONTINUOUS switch on EXTERN TIMER CONT CONTINUOUS switch off EXTERN approx OA AN pressure at pipette tip TIMING MON EXTERN mode FAF TIL INPUT IV TIMER CONT O ADOK EXTERN PEI bar pressure at pipette tip IV TIMING MON TMER mode TL INPUT IS PI TIMER CONT O bar EXTERN approx pressure at pipette tip IV
15. of three connectors on the front panel of the PWR 03D to the rear panel of the respective EPMS E housing see Figure 3 Figure 4 A POWER LED indicates that the PWR 03D is powered on see Figure 3 left Power switch voltage selector and fuse are located at the rear panel see Figure 3 right Note The chassis of the PWR 03D is connected to protective earth and it provides protective earth to the EPMS E housing if connected UN aC GEGN SUE He TER SLOW o VIBVAC w 13 S060 J i SS lt i ia Api electronic Gob CE D 71732 Toru Gerry wenn NOK AORC OCA iia OVATE OUTPUT A oJFu a H Y j y A AA A A AAA A pa npl Figure 3 Left PWR 03D front panel view Right PWR 03D rear panel view Note This power supply is intended to be used with npi EPMS E systems only version 2 4 page 5 PDES 01AM User Manual 2 7 System Grounding EPMS 07 EPMS 03 The 19 cabinet is grounded by the power cable through the ground pin of the mains connector protective earth In order to avoid ground loops the internal ground is isolated from the protective earth The internal ground is used on the BNC connectors or GROUND plugs of the modules that are inserted into the EPMS 07 housing The internal ground and mains ground protective earth can be connected by a wire using the ground plugs on the rear panel of the instrument It is not possible to predict whether measurements will be less or more noisy with the internal grou
16. pressure source is connected an OUT coupling where the injection pipette or the microJECT headstage respectively is connected and an EXHAUST coupling where a low retain pressure can be applied in order to avoid capillary effects at the tip of the injection pipette analogous to the retaining current used in iontophoretic systems This pressure must be generated by an additional pneumatic device Please contact npi electronic for details Systems with microJECT are additionally equipped with a connector for controlling the micro valve inside the microJECT The injection pressure is preset by a precision pressure regulator The pressure is displayed on an analog manometer Additional pneumatic devices tubes fittings connectors filters etc are available Please contact npi electronic for details A calibration procedure allowing quantitative drug application is described by Hofmeier and Lux 1981 see chapter 6 3 2 PDES 01AM Components The following items are shipped with the PDES 01AM system Y Pressure controller module for the EPMS 07 system Y User manual version 2 4 page 7 PDES 01AM User Manual Optional accessories microJECT pipette holder fast switch time lt 1 ms Tubing Ejection holder Remote push button External valve slow switch time gt 10 ms JEE eS 3 3 Description of the Front Panel O PDES 01AM noi O ON EJECT EXHAUST z O 1 o A 0 o TTL REMOTE TIMING EXTERNAL INPUT CON
17. ption Pressure ejection is a convenient method for applying both ionic and non ionic solutions from micropipettes This method is popular for two reasons First it can apply to almost all solutions Second the amount of solution ejected can be monitored by viewing the tip of the micropipette under a microscope np PDES units are designed for pressure ejection of drugs in physiological and pharmacological studies PDES systems can be equipped with high pressure outputs up to Pmax 4 bar 58 psi for drug application with fine tipped micropipettes or with low range pressure output Pmax 0 7 bar 10 psi for controlled droplet application from large tipped pipettes The PDES 01AM is a system with Pmax 4 bar 58 psi The device is designed as a module for the npi EPMS 07 system It comprises of one channel consisting of a precision pressure regulator an electronically controlled valve or a very fast micro valve located in the microJECT an analog timer and an analog pressure display Ejection of drugs can be activated manually or via a TTL input Mode of operation and timing is selected by controls at the front panel If the internal timer is used the eject time is preset by a control and the time base can be selected by a toggle switch The operational status is indicated by a red green LED The pressure connectors are also located at the front panel of the instrument The injection channel is equipped with an INPUT coupling where the
18. sion 2 4 page 9 PDES 01AM User Manual REMOTE CONTROL connector BNC connector for connecting a remote control e g a pushbutton REMOTE CONTROL 9 TTL INPUT BNC connector for connecting a TTL signal for EXTERN operation or for using the internal timer in TIMER operating mode see also 10 10 Operation mode switch Switch for setting the operation mode CONT The valve is continuously open TIMER The internal timer is used The timer can be started via a TTL HIGH signal at 9 or by pushing a pushbutton connected to 8 EXTERN The valve is open as long as the input signal at 9 is high or the pushbutton 1s pressed OFF The module is switch OFF the valve is closed LED 13 is also off OFF The PDES module is turned OFF the valve is closed LED 13 is also off The source pressure value is still shown at the manometer 1 but opening of the valve is disabled in OFF position see also chapter 4 11 x10s x1s x0 01s x0 01s switch Switch to set the time base of the timer ranges x10 s x1 s x0 1 s or x0 01 s 12 TIMER potentiometer Potentiometer to set the ejection time The total ejection time is the reading of this potentiometer multiplied by the reading of the time base switch 11 Note Due to tolerances of resistors and capacitors the time preset with the analog timer may vary up to 20 version 2 4 page 10 PDES 01AM User Manual 13 ON EJECT LED ON EJECT Indi
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