Molecular Devices Axoclamp 900A User manual

UserGuidefor
Axoclamp™900A
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Computer‐controlled
MicroelectrodeAmplifier
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MolecularDevicesCorporation
Sunnyvale,California94089
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PartNumber2500‐0179Rev.A
800.998.MATE | www.autom8.com | 650 University Ave # 5, Berkeley, CA 94710 USAt e l 5 1 0 . 8 4 5 . 6 2 8 3 | f a x 5 1 0 . 6 6 5 . 3 9 7 5 | e - m a i l i n f o @ a u t o m 8 . c o m

Copyright
©Copyright2006,MolecularDevicesCorporation.Allrightsreserved.Nopartof
thispublicationmaybereproduced,transmitted,transcribed,storedinaretrieval
system,ortranslatedintoanylanguageorcomputerlanguage,inanyformorby
anymeans,electronic,mechanical,magnetic,optical,chemical,manual,or
otherwise,withoutthepriorwrittenpermissionofMolecularDevicesCorporation,
1311OrleansDrive,Sunnyvale,California,94089,UnitedStatesofAmerica.
Trademarks
Axoclamp900Aisatrademark.DigidataandpCLAMPareregisteredtrademarks
ofMolecularDevicesCorporation.Allothertrademarksarethepropertyoftheir
respectiveowners.
TechnicalAssistance
Fortechnicalassistance,productcommentsandfeedback,orhelptoresolvea
problem,pleasevisit Molecular Devices Technical Support webpage at
www.moleculardevices.com/pages/supportoryoumayreachusbyphoneat
1‐800‐635‐5577(USonly);elsewhere,contactyourlocalrepresentative.
Disclaimer
MolecularDevicesCorporationreservestherighttochangeitsproductsand
servicesatanytimetoincorporatetechnologicaldevelopments.Thisuserguideis
subjecttochangewithoutnotice.
Althoughthisuserguidehasbeenpreparedwitheveryprecautiontoensure
accuracy,MolecularDevicesCorporationassumesnoliabilityforanyerrorsor
omissions,norforanydamagesresultingfromtheapplicationoruseofthis
information.
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Contents
TableofContents
Chapter1:Introduction.................................................................................................................... 1
Chapter2:InstallationandBasicOperation................................................................................ 5
Installation......................................................................................................................................5
CheckList ................................................................................................................................... 5
InstallingHardware .................................................................................................................. 6
InstallingtheAxoclamp900ACommander........................................................................... 6
Input/OutputConnections ...........................................................................................................8
FrontPanel.................................................................................................................................. 8
RearPanel ................................................................................................................................... 9
Overview ......................................................................................................................................11
Axoclamp900ACommanderInterfaceOverview.............................................................. 12
Toolbar ...................................................................................................................................... 12
Meters........................................................................................................................................ 13
ModeButtons ........................................................................................................................... 13
ModeTabs................................................................................................................................. 14
ScaledOutputTabs.................................................................................................................. 15
SettingParametersintheAxoclamp900ACommander.................................................... 15
ToolbarButtonsintheAxoclamp900ACommander......................................................... 18
CommunicationwiththeAxoclamp900A........................................................................... 18
ModelCells...................................................................................................................................19
TheCLAMP‐1UModelCell ................................................................................................... 19
TheMCO‐2UModelCell........................................................................................................ 21
InstallationofModelCells...................................................................................................... 21
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UserGuideforAxoclamp900A
PowerSupply...............................................................................................................................21
SupplyVoltageSelection........................................................................................................ 21
ChangingtheFuse................................................................................................................... 22
Glitches...................................................................................................................................... 22
GroundingandHum ..................................................................................................................23
Chapter3:SetupandTutorials..................................................................................................... 25
Checklist........................................................................................................................................25
Axoclamp900AandAxoclamp900ACommanderSetup..................................................... 26
Tutorial1:CurrentClamp(IC) ..................................................................................................28
Tutorial2:DiscontinuousCurrentClamp(DCC) ...................................................................34
Tutorial3:DiscontinuousSingle‐ElectrodeVoltageClamp(dSEVC).................................. 37
Tutorial4:Two‐ElectrodeVoltageClamp(TEVC) .................................................................41
I.TEVCUsingtheCLAMP‐1UModelCell.......................................................................... 41
II.TEVCUsingtheMCO‐2UOocyteModelCellandVG‐9ABathClamp ..................... 46
DCRestore................................................................................................................................49
Tutorial5:AutoModeSwitching..............................................................................................50
ExampleofAutoModeSwitching ........................................................................................ 51
SetupofAutoMode ................................................................................................................ 52
Tutorial6:High‐VoltageCurrentClamp(HVIC) ...................................................................55
Chapter4:GuidetoElectrophysiologicalRecording............................................................... 59
GeneralAdvice.............................................................................................................................59
ChamberDesign ...................................................................................................................... 59
Perfusion ................................................................................................................................... 60
MechanicalStability ................................................................................................................ 60
Optics......................................................................................................................................... 61
BathElectrode .......................................................................................................................... 61
InterfacingaComputer........................................................................................................... 62
Noise..............................................................................................................................................62
MeasurementofNoise ............................................................................................................ 62
SourcesofNoise....................................................................................................................... 63
BathErrorPotentials ...................................................................................................................65
(1)MinimizeRb......................................................................................................................... 66
(2)ClampVUsingaBathClamporVirtualGround
b........................................................ 67
(3)IndependentlyMeasureVandSubtractitfromtheCHANNEL1Potential
b........... 68
Summary................................................................................................................................... 70
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Contents
Headstages....................................................................................................................................70
HS‐9ASeries............................................................................................................................. 70
VG‐9ASeries ............................................................................................................................ 70
CurrentClampCircuit ............................................................................................................ 71
MountingtheHeadstage ........................................................................................................ 72
BathConnection....................................................................................................................... 72
StaticPrecautions..................................................................................................................... 72
AcousticPick‐up...................................................................................................................... 72
CompatibilityoftheHS‐2andHS‐2AwiththeAxoclamp900A...................................... 73
GeneralCharacteristics ........................................................................................................... 73
TheMeaningofH.................................................................................................................... 74
GuidelinesforSelectingaHeadstage.................................................................................... 75
HeadstageConnection ............................................................................................................ 77
Interchangeability.................................................................................................................... 77
Cleaning .................................................................................................................................... 77
InputLeakageCurrent............................................................................................................ 77
InputResistance....................................................................................................................... 78
Holders..........................................................................................................................................78
HolderDesign .......................................................................................................................... 78
OptionalAg/AgClPellets ....................................................................................................... 80
HolderUse................................................................................................................................80
HolderMaintenance................................................................................................................ 81
SharpMicroelectrodeRecording............................................................................................... 82
MicroelectrodeProperties....................................................................................................... 82
FillingSolutions ....................................................................................................................... 85
ImpalingCells .......................................................................................................................... 86
TheoryofRecordingModes....................................................................................................... 87
Current‐ClampMode.............................................................................................................. 87
DiscontinuousCurrentClamp(DCC)Mode ....................................................................... 88
DiscontinuousSingle‐ElectrodeVoltageClamp(dSEVC)Mode...................................... 91
NoiseinDCCanddSEVCModes ......................................................................................... 96
TEVCMode .............................................................................................................................. 98
Filters...........................................................................................................................................107
–3dBFrequency..................................................................................................................... 107
TypesofFilters....................................................................................................................... 108
Blanking ......................................................................................................................................110
CurrentMeasurement............................................................................................................... 111
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UserGuideforAxoclamp900A
Virtual‐GroundCurrentMeasurement ..................................................................................112
Chapter5:CommanderReference............................................................................................. 115
AlwaysOnTop ..........................................................................................................................115
BridgeBalance............................................................................................................................ 115
BridgeBalanceOverview ..................................................................................................... 116
BridgeBalanceintheBath.................................................................................................... 118
BridgeBalanceintheCell..................................................................................................... 118
Buzz .............................................................................................................................................119
Clear ............................................................................................................................................120
ConfigureSoftPanel ..................................................................................................................120
DefaultSoftPanelSettings .................................................................................................... 121
HowtoConfigureSoftPanel ................................................................................................ 121
DetectOscillation....................................................................................................................... 123
CurrentClamp ....................................................................................................................... 123
VoltageClamp........................................................................................................................ 124
CurrentInjectionResistor,R0................................................................................................... 124
Holding .......................................................................................................................................124
HowtoUseHolding ............................................................................................................. 125
KeyboardShortcuts................................................................................................................... 125
HotKeys.................................................................................................................................. 125
ShortcutKeys ......................................................................................................................... 125
LoadConfiguration...................................................................................................................128
ModeSwitching.........................................................................................................................129
ConnectinganExternalCommandSignal ......................................................................... 129
AutomaticModeSwitching.................................................................................................. 130
AdditionalNote ..................................................................................................................... 130
MonitorScope ............................................................................................................................131
Options........................................................................................................................................ 131
Options/General....................................................................................................................... 131
ExternalCommandSensitivity ............................................................................................ 132
LowpassFilterType .............................................................................................................. 133
SyncOutput............................................................................................................................ 133
AuxiliaryHeadstageInformation........................................................................................ 134
Miscellaneous......................................................................................................................... 134
Options/Auto............................................................................................................................ 135
SwitchtofromI‐ClamptodSEVC/TEVC......................................................................... 135
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