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Voltage clamping with single microelectrodes: Comparison of the discontinuous mode and continuous mode using the Axoclamp 2A amplifier

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Summary

The voltage clamp technique is a powerful method for studying the physiology of excitable membrane. This technique has made possible the determination of ionic responses generated by activation of either receptor-mediated or voltage-dependent processes. The development of the whole-cell, ‘tight-seal’ voltage clamp method has allowed the analysis and examination of membrane physiology at the single cell level. The method allows the characterization of voltage-dependent ionic conductances both at the macroscopic (whole-cell) and at the microscopic (unitary conductance or single channel) level in cells less than 10 µm in diameter, a feat difficult to achieve with ‘conventional’ fine-tipped micropipettes.

In this paper, several methologies used for culturing neuronal and non-neuronal cells in the laboratory are described. A comparison between the two modes of voltage clamp using blunt-tipped ‘patch’-microelectrodes, the switching (discontinuous) and the non-switching (continuous) modes, of the Axoclamp-2A amplifier is made. Some results on membrane currents obtained from neuronal and non-neuronal cells using the single electrode whole-cell ‘tight-seal’ voltage clamp is illustrated. The possible existence of two inactivating K+ currents, one dependent on Ca++ the other is not, is discussed.

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  1. Department of Physiology and Biophysics, University of Cincinnati, College of Medicine, 45267-0576, Cincinnati, OH, USA

    R. Y. K. Pun

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Pun, R.Y.K. Voltage clamping with single microelectrodes: Comparison of the discontinuous mode and continuous mode using the Axoclamp 2A amplifier. Mol Cell Biochem 80, 109–120 (1989). https://doi.org/10.1007/BF00231009

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