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Perforated Whole-Cell Patch-Clamp Technique: A User’s Guide

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Patch Clamp Techniques

Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

The patch-clamp technique has revolutionized the study of membrane physiology, enabling unprecedented resolution in recording cellular electrical responses and underlying mechanisms. The perforated-patch variant of whole-cell patch-clamp recording was developed to overcome the dialysis of cytoplasmic constituents that occurs with traditional whole-cell recording. With perforated-patch recordings, perforants, such as the antibiotics nystatin and gramicidin, are included in the pipette solution and form small pores in the membrane attached to the patch pipette. These pores allow certain monovalent ions to permeate, enabling electrical access to the cell interior, but prevent the dialysis of larger molecules and other ions. In this review we give a brief overview of the key features of some of the perforants, present some practical approaches to the use of the perforated patch-clamp mode of whole-cell (PPWC) recordings, and give some typical examples of neuronal responses obtained with the PPWC recording that highlight its utility as compared to the traditional whole-cell patch recording configuration.

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Authors and Affiliations

  1. Department of Developmental Physiology, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki, Aichi, Japan

    Hitoshi Ishibashi & Junichi Nabekura

  2. Department of Physiology, School of Medical Sciences, University of New South Wales, Sydney, Australia

    Andrew J. Moorhouse

Authors
  1. Hitoshi Ishibashi
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  2. Andrew J. Moorhouse
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  3. Junichi Nabekura
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Corresponding author

Correspondence to Hitoshi Ishibashi .

Editor information

Editors and Affiliations

  1. National Institute for Physiological Sciences National Institutes of Natural Sciences, Okazaki, Aichi, Japan

    Yasunobu Okada

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© 2012 Springer

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Ishibashi, H., Moorhouse, A.J., Nabekura, J. (2012). Perforated Whole-Cell Patch-Clamp Technique: A User’s Guide. In: Okada, Y. (eds) Patch Clamp Techniques. Springer Protocols Handbooks. Springer, Tokyo. https://doi.org/10.1007/978-4-431-53993-3_4

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  • DOI: https://doi.org/10.1007/978-4-431-53993-3_4

  • Publisher Name: Springer, Tokyo

  • Print ISBN: 978-4-431-53992-6

  • Online ISBN: 978-4-431-53993-3

  • eBook Packages: Springer Protocols

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