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PC-505B Manual, Rev. 201013

Warner Instruments

Whole Cell/Patch Clamp Amplifier

Model PC-505B

Warner Instruments

1125 Dixwell Avenue, Hamden, CT 06514

(800) 232-2380 / (800) 547-6766 - support

www.warneronline.com

Summary of Contents for PC-505B

Page 1: ...C 505B Manual Rev 201013 Warner Instruments Whole Cell Patch Clamp Amplifier Model PC 505B Warner Instruments 1125 Dixwell Avenue Hamden CT 06514 800 232 2380 800 547 6766 support www warneronline com...

Page 2: ...mand block 11 METER section 13 Front and rear panel BNC s and connectors 13 Headstage 16 Electrode holders 16 Model Cell 17 SETUP AND INITIAL TEST 18 Line voltage 18 Instrument grounding 18 Headstage...

Page 3: ...ng Cell attached patch 32 Whole cell recording Initial conditions 32 Whole cell recording Rupture the membrane patch 32 Whole cell recording Series R compensation 33 Whole cell recording correction 34...

Page 4: ...PC 505B Manual Rev 201013 Recommended reading 44 Glossary 47...

Page 5: ...at 1 kHz Built in RMS noise monitor Voltage and current clamp modes with independently selectable Vhold and Ihold Bandwidth to 25 kHz Internal 4 pole low pass Bessel filter with rear panel filter tele...

Page 6: ...italic type Special comments and warnings are presented in highlighted text Any other formatting should be apparent from context Device panel abbreviations Many controls on the PC 505B have abbreviati...

Page 7: ...ell attached patches the physiological transmembrane current equals minus the indicated Im Membrane potential Vm Membrane potential is defined as pipet potential minus bath potential With outside out...

Page 8: ...and 1 0 nA respectively A MODE toggle switch in the OUTPUT command block is used to select between voltage clamp Vc or current clamp Ic modes Placing the instrument in current or voltage clamp mode ac...

Page 9: ...COMMAND IN BNC are attenuated and available to be applied to the headstage Junction Zero and Auto Zero These controls set the pipette current to zero after the pipette is placed in the test solution...

Page 10: ...membrane to the command signal waveform and 2 to minimize large current transients associated with rapid changes in applied membrane potential These effects if uncompensated can drive the headstage am...

Page 11: ...bath through the pipet membrane seal resistance RS or through the membrane patch For leaky seals RS 1 G it is important to correct for the error caused by the shunt resistance of the leak This error c...

Page 12: ...A with the 50 M resistor NOTE Current clamp mode does not operate with the PROBE SELECT toggle set to high The low headstage resistance is automatically selected when in current clamp mode Gain The GA...

Page 13: ...V Vm This selection reports the transmembrane potential when in current clamp mode Full scale range is 200 mV Im This selection reports the transmembrane current when in voltage clamp mode Full scale...

Page 14: ...ince the center pin and sleeve are the and inputs respectively the sleeve is not grounded The applied voltage is attenuated by the COMMAND SENSITIVITY switch and is available in both voltage and curre...

Page 15: ...ers and selector function switches for PSTAT and SPEED TEST modes Headstage cable connector The headstage connects to the instrument via an 8 pin DIN connector PSTAT switch The PC 505B is converted to...

Page 16: ...ich is driven by the command signal The attached metal rod is used for mounting to a micromanipulator and is insulated from the headstage housing A 2 5 m cable with 8 pin DIN connector attaches the he...

Page 17: ...rovided for applying suction to seal the pipet tip to the cell membrane NOTE The standard holder used with the PC 505B is type QSW AxxP straight body where xx specifies the glass OD in mm Model Cell A...

Page 18: ...video monitor walking across a dry carpet wearing polyester clothing etc to alter your electric potential by as much as 10 kV relative to ground A discharge of this magnitude through the headstage can...

Page 19: ...lifier The headstage mounted into a shielded enclosure An oscilloscope with BNC cables An adjustable DC voltage source The model cell shipped with the instrument Initial connections Place the amplifie...

Page 20: ...ANDS off ZAP toggle COMMANDS safe ZAP control COMMANDS 0 1 ms C FAST 1 FAST CAP COMP fully CCW C FAST 2 FAST CAP COMP fully CCW FAST 1 FAST CAP COMP fully CCW FAST 2 FAST CAP COMP fully CCW C SLOW WHO...

Page 21: ...h to the Junction Zero 2 Adjust the JUNCTION ZERO control COMMANDS block until the meter reading is 10 mV 3 Verify that Vm and Vc on the METER SELECTOR switch give the same meter reading as Junction Z...

Page 22: ...mV 17 Set METER SELECT to Vc Verify that meter reads sum of holding potential and junction offset 250 mV 18 Set METER SELECT control to Vc h IN Verify that the adjustment of JUNCTION ZERO control does...

Page 23: ...nnect voltage source from COMMAND IN BNC 2 Connect model cell to headstage in patch mode 3 Set METER SELECT to Im 4 Set PROBE RESISTER to high 5 Set COMMAND SENSITIVITY toggle to off 6 Set COMMAND SEL...

Page 24: ...Leading edge transients will remain due to characteristics of model cell 9 Set PROBE RESISTER to low 10 Connect model cell to headstage in whole cell mode 11 Turn C SLOW toggle switch on 12 Adjust C...

Page 25: ...is disabled in Io mode 10 Set MODE toggle to Ic 11 Set METER SELECT to Im 12 Adjust CURRENT HOLD control until METER reads 500 pA 13 Set METER SELECT to Vm 14 Verify that meter reads 5 1 mV correspond...

Page 26: ...OMMANDS fully CCW toggle switch off CURRENT HOLD COMMANDS fully CCW toggle switch off COMMAND SELECT COMMANDS external command COMMAND SENSITIVITY COMMANDS off x0 001 JUNCTION ZERO control COMMANDS mi...

Page 27: ...y delay will foul the tip and inhibit seal formation Also it is advisable to locate the cell you want to patch before lowering the tip into solution Just before lowering the electrode pipette tip into...

Page 28: ...sure the reference electrode is properly submerged If the zero setting continues to drift then check your setup for errors e g chloriding of the electrodes IMPORTANT Once set do not make further adjus...

Page 29: ...e the pipette begins to dimple the cell membrane the amplitude of the test current should begin to decrease When this happens release the applied positive air pressure in the electrode holder and watc...

Page 30: ...nks of CAP COMP controls 1 2 interact with each other some trial and error in making a good adjustment is required Compensation is optimal when the square wave has as sharp a rise and fall as possible...

Page 31: ...RACTION control clockwise until the Im trace on your acquisition system or oscilloscope becomes flat except for residual capacitance transients Alternatively with the TEST PULSE off run the holding po...

Page 32: ...to a fixed value before beginning to record or to reset it to another fixed value at any time during an experiment first set the meter to Vc h IN then set the COMMANDS VOLTAGE POLARITY toggle switch t...

Page 33: ...ll slow the response of the amplifer Series R compensation is adjusted by using the SERIES R and C SLOW controls The SERIES R control compensates for the resistance of the pipette opening at the cell...

Page 34: ...ng correction Once the series resistance has been measured and corrected you can further optimize the amplifier by applying a signal boost in the form of correction Generally speaking the maximum corr...

Page 35: ...om those made while in voltage clamp mode They are still valid when switching back to voltage clamp mode Changes made now can t be monitored can result in over compensation feedback oscillation and ca...

Page 36: ...ge The HB 205B is a switchable 50 G 500 M headstage modified to allow for compensating the relatively large capacitance of the bilayer membrane Up to approximately 250 pF can be compensated As such th...

Page 37: ...rbonate holder body as this can weaken the structure Chloriding by electrolysis Dip the Ag wire to the required depth in a solution of 0 1M NaCl or KCl optionally made slightly acidic with HCl and arr...

Page 38: ...r rinse thoroughly with distilled water dry thoroughly and reassemble Cleaning and storage After use rinse any deposits from the holder body the Ag wire pin jack and suction port and allow to dry To p...

Page 39: ...he Ag AgCl coating can be submerged with no bare Ag exposed to solution The free end is connected to circuit ground Any exposed Ag surface that could potentially contact the bathing solution should be...

Page 40: ...V with 10 turn control Internal Test Pulse 1 Volt 50 60 Hz line frequency square wave attenuated by COMMAND SENSITIVITY Command Sensitivity attenuates at x0 1 x0 01 and x0 001 Zap 1 0 Volt Pulse adjus...

Page 41: ...ommands 199 9 mV Vm membrane voltage in current clamp mode 199 9 mV Im membrane current 1999 pA RMS noise 1 999 pA REAR PANEL OUTPUTS Gain Telegraphs 0 5 to 7 0 V in 0 5 V steps Filter Telegraphs 0 2...

Page 42: ...rs Lengths up to 3 05 meters 10 are available Electrode Holders One electrode holder is used with each headstage The standard model is QSW AxxP straight Holder part numbers are completed by replacing...

Page 43: ...place This warranty is not extended to electrode holders since these items are considered disposable Service notes A If the instrument POWER light fails to light check the fuse at the rear panel If th...

Page 44: ...subject to unnecessary expense in clearing U S Customs Recommended reading Sakmann and Neher 1983 Single channel Recording Plenum Press Hamill Marty Neher Sakmann and Sigworth 1981 Pflugers Archiv 391...

Page 45: ...0 4 2 EN61000 4 3 ENV50204 EN610000 4 4 EN610000 4 8 EN610000 4 11 Manufacturer s Name Warner Instruments LLC Manufacturer s Address 1125 Dixwell Avenue Hamden CT 06514 Tel 203 776 0664 Equipment Desc...

Page 46: ...ts LLC Manufacturer s Address 1125 Dixwell Avenue Hamden CT 06514 Tel 203 776 0664 Equipment Description Instrument Amplifier Safety requirements for electrical equipment for measurement and laborator...

Page 47: ...PC 505B Manual Rev 201013 Warner Instruments A Harvard Apparatus Company 47...

Page 48: ...pacitance A capacitor can be represented by a small break in a conducting pathway bounded by two parallel plates The electric field generated across the space between the plates in the presence of an...

Page 49: ...gs with the same outer diameter and appearance electrode One terminal of a voltage source which can either supply or collect current electromagnetic From physics An electric current induces a magnetic...

Page 50: ...t which repeats itself at regular intervals perfusate The solution being perfused perfusion The exchange of one solution with another plasma membrane The surface membrane of a cell Contrast with an in...

Page 51: ...Determined by measuring the current through an open channel in the presence of a driving force transmembrane potential at different potentials Measurements made within the Ohmic range of the channel s...

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