The extracellular patch clamp: a method for resolving currents through individual open channels in biological membranes

Pflugers Arch. 1978 Jul 18;375(2):219-28. doi: 10.1007/BF00584247.

Abstract

The current contributions of individual ionic channels can be measured by electrically isolating a small patch of membrane. To do this, the tip of a small pipette is brought into close contact with an enzymatically cleaned membrane of a hypersensitive amphibian or mammalian muscle fiber. Current flowing through the pipette is measured. If the pipette contains cholinergic agonist at mu-molar concentrations, square pulse current waveforms can be observed which represent the activation of individual acetylcholine-receptor channels. The square pulses have amplitudes of 1 to 3 pA and durations of 10--100ms. In order to obtain the necessary resolution, a delicate compromise had to be found between different experimental parameters. Pipettes with 1--3 micrometer internal diameter and a steep final taper had to be used, extensive enzyme treatment was necessary, and conditions had be to found in which channels open at a relatively low frequency.

MeSH terms

  • Animals
  • Anura
  • Cell Membrane / metabolism
  • Cell Membrane / physiology*
  • Electric Conductivity
  • Ion Channels*
  • Muscles / metabolism
  • Muscles / physiology
  • Physiology / instrumentation
  • Physiology / methods*
  • Rats
  • Receptors, Cholinergic / physiology

Substances

  • Ion Channels
  • Receptors, Cholinergic