A novel way to go whole-cell in patch-clamp experiments

IEEE Trans Biomed Eng. 2010 Nov;57(11). doi: 10.1109/TBME.2010.2055055. Epub 2010 Jun 28.

Abstract

With a conventional patch-clamp electrode, an Ag/AgCl wire sits stationary inside the pipette. To move from the gigaseal cell-attached configuration to whole-cell recording, suction is applied inside the pipette. We have designed and developed a novel Pushpen patch-clamp electrode, in which a W wire insulated and wound with Ag/AgCl wire can move linearly inside the pipette. The W wire has a conical tip, which can protrude from the pipette tip like a push pen, a procedure we call the Pushpen Operation. We use the Pushpen operation to impale the cell membrane in cell-attached configuration to go whole-cell without disruption of the gigaseal. We successfully recorded whole-cell currents from chinese hamster ovarian cells expressing influenza A virus protein A/M2, after obtaining whole-cell configuration with the Pushpen operation. This novel method of achieving whole-cell configuration may have a higher success rate than is the case with the conventional patch clamp technique.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • CHO Cells
  • Cell Membrane / chemistry
  • Cell Membrane / metabolism
  • Cell Membrane / physiology
  • Cricetulus
  • Electrophysiology / instrumentation*
  • Electrophysiology / methods
  • Equipment Design
  • Humans
  • Patch-Clamp Techniques / instrumentation*
  • Patch-Clamp Techniques / methods
  • Viral Matrix Proteins / metabolism

Substances

  • M2 protein, Influenza A virus
  • Viral Matrix Proteins