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Determination and compensation of series resistances during whole-cell patch-clamp recordings using an active bridge circuit and the phase-sensitive technique

  • Neuroscience
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Abstract

We present a technique which combines two methods in order to measure the series resistance (R S) during whole-cell patch-clamp recordings from excitable and non-excitable cells. R S is determined in the amplifier’s current-clamp mode by means of an active bridge circuit. The correct neutralization of the electrode capacitance and the adjustment of the bridge circuit is achieved by the so-called phase-sensitive method: Short sine wave currents with frequencies between 3 and 7 kHz are injected into the cells. Complete capacitance neutralization is indicated by the disappearance of the phase lag between current and voltage, and correct bridge balance is indicated by a minimized voltage response to the sine wave current. The R S value determined in the current-clamp mode then provides the basis for R S compensation in the voltage-clamp recording mode. The accuracy of the procedure has been confirmed on single-compartment cell models where the error amounted to 2–3 %. Similar errors were observed during dual patch clamp recordings from single neocortical layer 5 pyramidal cells where one electrode was connected to the bridge amplifier and the other one to a time-sharing, single-electrode current- and voltage-clamp amplifier with negligible R S. The technique presented here allows R S compensation for up to 80–90 %, even in cells with low input resistances (e.g., astrocytes). In addition, the present study underlines the importance of correct R S compensation by showing that significant series resistances directly affect the determination of membrane conductances as well as the kinetic properties of spontaneous synaptic currents with small amplitudes.

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Acknowledgments

We thank F. Rucker for mathematical support, G. Horn for technical assistance, R. Kurtz for valuable comments on the manuscript, and M. Sutor-Gauß for editing the manuscript.

BS would like to dedicate this manuscript to the 80th birthday of Prof. Dr. Gerrit ten Bruggencate.

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Authors and Affiliations

  1. BioMedizinisches Centrum (BMC), Ludwig-Maximilians-University (LMU), Physiological Genomics, Physiological Institute, Großhaderner Str. 9, 82152, Planegg, Germany

    Therese Riedemann & Bernd Sutor

  2. npi electronic GmbH, Bauhofring 16, 71732, Tamm, Germany

    Hans Reiner Polder

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  1. Therese Riedemann
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  2. Hans Reiner Polder
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  3. Bernd Sutor
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Correspondence to Bernd Sutor.

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HRP is an employee of npi electronic GmbH. TR and BS declare no conflicts of interest.

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Riedemann, T., Polder, H.R. & Sutor, B. Determination and compensation of series resistances during whole-cell patch-clamp recordings using an active bridge circuit and the phase-sensitive technique. Pflugers Arch - Eur J Physiol 468, 1725–1740 (2016). https://doi.org/10.1007/s00424-016-1868-8

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  • DOI: https://doi.org/10.1007/s00424-016-1868-8

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