pulse and pulsefit

E l e c t r o p h y s i o l o g y E l e c t r o c h e m i s t r y

HEKA provides the finest instruments today to achieve the needed progress of tomorrow…

PULSE / PULSEFIT

Software for electrophysiological research,data acquisition, review and online analysis

Features PULSEFIT

• Data Review

• Cursor-operated measuring

Features

• Fit of Data Traces

• Fit of Parameters

PULSE

• Digital Oscilloscope

• Versatile Pulse Generator

• Parameter Monitoring

• Data Tree Editor

• Online Analysis

PULSE is easily adaptable to

any individual approach for

performing electrophysio-

logical experiments.

P U L S E / P U L S E F I T

PULSE and PULSEFIT - a versatile program for physiological research

PULSE/PULSEFIT - a program

which first thinks of data

analysis

The consideration of all meas-uring parameters has madePULSE the most powerfulprogram for electrophysiologicalclamp experiments. It has beendeveloped with the help ofleading laboratories worldwideto meet the requirements of thescientific community. PULSEand PULSEFIT thus providesthe optimal solution for dataacquisition and evaluation.Further programs such as TACand PULSETOOLS can imme-diately process the data ob-tained with PULSE. Exportfunctions transfer the data toother programs such as Igor, oras ASCII (MAC and DOS). The"extensions" of PULSE(LOCK-IN, FURA, and X-CHART)provide the link from clamp tofurther experiments.

PULSE/PULSEFIT - a reliable

and convenient program

PULSE and PULSEFIT consti-tute the perfect combination ofsimple operation and com-prehensive functionality. For thebeginner, the operation can belimited to the essential func-tions. Intuitive operation anduncompromising simplificationof complicated adjustmentsquickly enable the user toperform more complexexperiments and evaluations.The experienced patch-clampinvestigator will be fascinatedby the versatility of the programand logical structure of thefunctions. The outstanding userinterface of the program with"virtual control keys" can bedefined by the user to meet hisrequirements. From the veryfirst day you can record datafrom your experiments withPULSE.PULSE and PULSEFIT providethe optimum in userconvenience.

PULSE/PULSEFIT - a program

for your amplifier, too

PULSE is custom-made forapplication with the EPC 9 andEPC 10 patch clamp amplifiers,but any other amplifier also re-ceives a tremendous boost withPULSE, which supports allpatch-clamp and voltage-clampamplifiers. Therefore, you canoperate various set-ups withdifferent amplifiers and pur-poses in a laboratory with thesame software.All data are available in thesame documented format, andyou can thus prepare filters foryour own programs. Do youhave questions? If so, a hotlineis at your disposal!

Picture front page:Purkinje neuron and Bergmann glialcell in a cerebellar slice preperation,T. Möller, MDC, Berlin


PULSE provides sophisticated tools for electrophysiological experiments

Replay

Acquired data and old data filescan be reviewed and edited.

Online Analysis

Immediate analysis of the justacquired or replayed data.The analysis result is shown asa function of a variety of para-meters, specified by the user.

Parameters

Parameters allow the additionaldata acquisition of differentvalues as RMS noise, pipetteresistance, seal resistanceor user defined parameters(e.g. pH). Functions

Function features allow thedisplaying of measuredresponses like leak current…

Sequence Pool

Pool of available pulse templates.Up to six template controls aredisplayed at a time, which canbe stopped or interrupted by thetools.

Notebook

During stimulation and replaythe results of the online analysisare displayed in the notebook.These data columns can beexported to disk or copied tothe clipboard.

Tree

Export your data (e.g. singlesweep, online analysis results)to printer, IGOR files (presentingand analysing program) or ASCII(MAC or DOS).

Buffer

Add, substract or accumulateyour measured sweeps.

Marks

Tag several entries in the datafile to perform one action to allof them (e.g. print, average).

Display

Change the display of theoscilloscope(e.g. grid, dimmed overlay).


EPC 9 - the virtual front panel

The Pulse Generator window provides the layout of a stimulation pulse sequence

Test Pulse

There are two test pulse modes: built-in testpulses (double or single) and use of a stimulationtemplate from the pulse generator as test pulse.

Leak and Rs Compensation

This controls a hardware leak compensation. Theseries resistance compensation corrects formembrane voltage errors under conditions of highaccess resistance between pipette and cellinterior.

Stimulus

The stimulus can be filtered by activating thestimulus switch.

Automatic Compensations

Automatic routines for leak and capacitivetransient compensations, perform these tasksfaster and more accurate than even the mostexperienced experimentor. Capacitance trackingallows continuous updating of membranecapacitance and series resistance compensationduring recording sessions.

Documentation

All EPC 9 settings are stored with the raw datafor easy reconstruction, analysis, and docu-mentation of your experimental procedures.

Filters

Two built-in high quality hardware filters(Butterworth/Bessel) perform excellent signalconditioning and remove the expense ofpurchasing additional filter instruments.

Controls

All values can be set automatically or manuallyby either the mouse or the keyboard.

V-Membrane

Displays the pre-sently selectedholding potential.

Leak

Determines diffe-rent parameters ofthe leak pulses.

Timing

Determines thenumber of thesweeps, thesweep- and thesample interval.

Segments

A pulse patternconsists of anarbitrary numberof segments.

AD/DA Channels

Switch betweendifferent triggermodes and choosethe right channelsfor acquisition.

Increment Mode

The variable deter-mines the order ofincrement of thesegment voltage/duration.

Triggers

There are up to 3triggers chooseable.

Sequence Pool

It is a paging bar with a nearly unlimited numberof sequences. Loads, saves, copies etc. the poolof available stimulation sequences.

Zap Pulse

A high voltage pulse is applied to the pipette inorder to rupture the patch membrane. The para-meters (duration and amplitude) can be specified.

Macros

Macro features allow the recording of routinefunctions and then accessing these Macros by asimple mouse click.

Sequence

Cartoon

After each editing operation the pulse pattern isdisplayed in the cartoon.


PULSEFIT - all common fit routines

in the field of electrophysiology are available

Sweep Fit

The trace fit is superimposedto the raw data within a win-dow selected by the verticalcursor lines. In this case, adouble-exponential fitdescribes the current decay.

Simplex Fit

Control parameters for theSimplex fit algorithm are editedhere. Manual fit can beperformed for the determinationof good starting values.

Sweep Fit Control

Buttons for the control of fits toraw data traces. “Auto” repeatsthe same kind of Sweep Fit for allSweeps of a series automatically.

Fit Type

Select a fit function (polynomial,Gaussian, function according toa Hodgkin & Huxley formalismfor channel activation and in-activation) or store results ofthe Online Analysis or thoseobtained from manual mouse-operated measurements tothe Analysis file.

Fit Function parameters

The parameters of the selectedfit function are displayed. In thiscase, up to three exponentialscan be fitted.

Sweep Fit Results Table:

The results determined bySweep Fit are stored in theAnalysis File and are available inSeries Fit for further analysis.These parameters can bedisplayed as a function of otherparameters or various abscissavariables like potential orduration of the relevantsegment.

Series Fit Control

Control buttons for Series Fit.Selected data and fits can beexported in various formats.Individual data points can beskipped so they are notconsidered for the fit.

Fit Types

Fit functions supported by Series Fit are: Polynomials, Expo-nentials, Boltzmann functions, and current-voltage relation-ships. In this example recovery from inactivation is analyzed.The peak current in a test segment is plotted versus the expo-nentially increasing duration of an interpulse segment. Thetheoretical curve describes a double exponential recovery.

The PULSE software and ourEPC 9 or EPC 10 patch clampamplifier combine to provide youwith everything you need if youare interested in membranecapaci-tance measurements.The EPC 9 or EPC 10 is the idealpatch clamp amplifier forcapacitance measurementsbecause all the relevantparameters are under its control.Digital control of filter settings,gain and compensation networksallows the admittance of thesignal source at the amplifier'sinput to be determined based onthe amplifier's own calibration.No external filters are requiredfor standard measure-ments.

In 1982 Neher and Marty in-troduced the lock-in amplifier intothe patch clamp field for mea-surement of membrane capa-citance using a single sine-wavefrequency. For determining theappropriate phase setting, theyused a simple and easy trick -dithering of the compensation

Capacitance measurements with PULSE

implemented in the PULSEsoftware together with twofurther variants of capacitancemeasurement.

The PULSE software implementsboth the classical singlefrequency and the 1 1/2frequency (or Lindau-Neher)technique in a convenient user-friendly way. It provides 'internalcalibration' which automaticallycorrects for phase-shifts andfrequency dependent attenuation,so that the admittance of theexternal circuit is obtained incalibrated SI units. On this basisthe parameters of the equivalentcircuit are calculated 'online'according to Lindau & Neher.

network while changing thephase for obtaining a maximumsignal. This method is appropriateunder stationary conditions.

Commonly used extensions,however, particularly the auto-matic 'phase tracking', is proneto errors (see Gillis in B. Sakmann& E. Neher Eds. Single ChannelRecording 2nd Edition, Plenum).For this reason several novelschemes, including 2 frequencymethods, have been suggestedin the meantime. The minimumrequirement for determining thethree unknowns of the equivalentcircuit of the cell under studyinvolves three experimentalparameters. The 1 1/2 frequencymethod (or Lindau-Neher tech-nique) uses real and imaginarypart of a sine-wave signal plusthe DC-conductance to deter-mine membrane capacitance,membrane conductance, andaccess resistance. This techniquewas adopted long ago by leadingpatch clamp laboratories. It is fully

In addition, the EPC 9 or EPC 10offers automatic capacitancecompensation. When activatedrepetitively (capacitance tracking!)this provides a continuousreadout of capacitance. Theexperimentor can observe theresidual capacitive current for avalidation of correctcompensation.

Since the amplifier 'knows' thesettings of the compensationnetwork, the 1 1/2 frequencymethod can operate while thebulk of the capacitance iscompensated. Consequently theamplifier can be operated at highgain without saturating. High gainimplies low noise. The highresolution admittancemeasurement of the residualcapacitance together with thecompensation circuit provides aneffective dynamic range whichhas yet to be achieved with anyother instrument.



Service & Support

As the first manufacturer ofpatch clamp amplifiers in theworld HEKA knows the needsof scientists. We provideexceptional pre and post salescustomer support from ourtrained international sales re-presentatives and our owntechnical support advisors. Withthousands of high performancehardware and software prod-ucts in daily use worldwide weunderstand all aspects of dataacquisition systems not just thesoftware.

Related Products

X-CHART

Software implementedchart-recorder.FURA

Extension for synchronizedratiometric fluorescencemeasurementsPULSETOOLS

Software for data reorganisation,editing and averaging. Amplitudehistograms and noise analysis.PULSESIM

Modeling and simulation of databased on kinetic schemes ordiscrete Markov models.EPC 10

The fully computer controlledpatch clamp amplifier withbuilt-in interface board.LIH 1600

16 bit multi-channel dataacquisition systemEPC 8

The successor of the EPC 7.Manual or digital controlselectable.EPC 7 PLUS

The classic patch clampamplifier for single channel andwhole-cell measurements.PIP 5

Temperature controlled micropipette puller.

Technical specifications

PULSE

Data acquisition• 2 channels A/D and 1

channel D/A (stimulation)• 3 channels D/A (Trigger,

auxiliary stimulation channels)

Amplifier

All 'clamp' amplifiers aresupported. Several amplifierwindows for e.g. EPC 9,EPC 10 and TelegraphingAmplifiers are available.Other amplifier windows canbe generated.Extensions to Pulse

• X-CHARTMultichannel chart recorder for up to 16 channels.

• FURAPerforms synchronized ratiometric fluorescence- andelectrophysiological measurements.

• LOCK-INSoftware lock-in to measure membrane capacitance.

PULSEFIT

PULSEFIT is an analysisprogram for pulsed data files.Data can be analyzed on threehierarchical levels:Sweep FitFit of raw data traces. Withinspecified windows variousfunctions can be fitted with aSimplex algorithm to the rawdata traces:Raw Data Fits• Polynomials• Exponentials• Gaussian Fit• Versatile functions according

to Hodgkin & Huxley gatingformalism

Series FitFit of parameters as derivedfrom the fit to raw traces (e.g.peak current) on the level of aseries of traces as a function ofvarious parameters (e.g. pulse,potential):Result Fits• Polynomials• Exponentials• Boltzmann Laws• Various Current-Voltage

RelationshipsGroup FitFit of parameters as derivedfrom the fit to data on the levelof a series (e.g. reversal poten-tials) as a function of variousparameters (e.g. concentrationof an ingredient of the bathingsaline):Parameter Fits• Polynomials• Exponentials• Boltzmann Laws• Dose-Response Curves

PULSE/PULSEFIT

In/Out & data management• Export to ASCII (MAC and

DOS) format), IGOR• Printout of raw data and

Online Analysis resultsGraphical representation• Oscilloscope• Zoom• Dimmed Overlay• Absolute and relative Cursor

MeasurementsProcessing• p/n Leak Correction• Zero Correction• Software Gaussian Filter• Buffer (basic arithmetic

Operation)• Fit of Data TracesMiscellaneous• Runs on Mac OS up to

version 9 and Windows 98/NT/2000/XP

• Macro Recording• Online Analysis

Hardware requirements

• For Data acquisition EPC 10 or EPC 9 amplifier

or LIH 1600, ITC-16 or ITC-18 acquisition interface

• ComputerSee data sheet “Recommended Configuration”

VK

RP

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General notice:

Product names used hereinare for identification purposesonly and may be trademarksof their respective owners.HEKA disclaims any and allrights in those marks.

We reserve the right to effecttechnical changes as develop-ment progresses.Special versions are availableon request. Further technicaldata are provided by a de-tailed description, which isavailable on request.A guarantee of one yearapplies on all instruments.

HEKA Instruments Inc.33 Valley RoadSouthboro, MA 01772USA

+1 866 742 0606 (Toll Free)+1 508 481 8945http://[email protected]@heka.com

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HEKA Electronics Incorporated47 Keddy Bridge RoadR.R. #2Mahone Bay, NS B0J 2E0Canada


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+49 (0) 63 25 / 95 53-0+49 (0) 63 25 / 95 53-50http://[email protected]@heka.com

pulse and pulsefit

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