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Perforated Whole-Cell Recordings in Automated Patch Clamp Electrophysiology

Part of the Methods in Molecular Biology book series (MIMB,volume 2188)

Abstract

The automated patch clamp (APC) technology is used for increasing the data throughput of electrophysiological measurements, especially in safety pharmacology and drug discovery. Typically, electrical access to the cells are obtained using standard whole-cell formation by rupturing the membrane, thereby causing a rapid washout of cytosolic components. In contrast the perforated whole-cell configuration provides electrical access to the cell interior while limiting intracellular wash-out. This method allows for recordings of ion channels that are gated by intracellular modulators (e.g., ATP, cyclic nucleotides, or Ca2+), prevents channel current “run down,” and maintains a physiological membrane potential for action potential recordings. Here we present some practical approaches to the use of perforated patch clamp for APC recordings. Our findings from these high-throughput, data-rich measurements (e.g., defining optimized concentrations and practical recommendations for four different perforating agents) can be more broadly applied to perforated patch clamp experiments in general (automated and manual), improving success rates, experimental conditions, and applications.

Key words

  • Automated patch clamp
  • Perforated patch clamp
  • Perforating agent
  • Pore-forming agent
  • Nystatin
  • Amphotericin B
  • β-Escin
  • Gramicidin
  • Qube 384

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  • DOI: 10.1007/978-1-0716-0818-0_5
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Correspondence to Kadla R. Rosholm .

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Rosholm, K.R., Boddum, K., Lindquist, A. (2021). Perforated Whole-Cell Recordings in Automated Patch Clamp Electrophysiology. In: Dallas, M., Bell, D. (eds) Patch Clamp Electrophysiology. Methods in Molecular Biology, vol 2188. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0818-0_5

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  • DOI: https://doi.org/10.1007/978-1-0716-0818-0_5

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0817-3

  • Online ISBN: 978-1-0716-0818-0

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