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Related Concept Videos

Patch Clamp01:18

Patch Clamp

Many fundamental cell functions such as muscle contraction and nerve transmission rely on the electrical signals produced by the movement of positively and negatively charged ions across the cell membrane. One competent method to record current flowing across the whole cell or single ion channel is the patch-clamp technique.
In this method, a glass micropipette containing electrolyte solution is tightly sealed against a small portion of the cell membrane. As a result, a patch of the cell...

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Culturing and Electrophysiology of Cells on NRCC Patch-clamp Chips
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Published on: February 7, 2012

Robotic multiwell planar patch-clamp for native and primary mammalian cells.

Carol J Milligan1, Jing Li, Piruthivi Sukumar

  • 1Institute of Membrane and Systems Biology, Faculty of Biological Sciences, Garstang Building, University of Leeds, Leeds LS29JT, UK.

Nature Protocols
|February 7, 2009
PubMed
Summary

Robotic multiwell planar patch-clamp technology enables efficient intracellular solution exchange for ion channel research. This method expands its utility beyond drug development to diverse cell types, including those from the nervous and immune systems.

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Area of Science:

  • Electrophysiology
  • Ion Channel Biology
  • Cellular Neuroscience

Background:

  • Robotic multiwell planar patch-clamp is widely used for drug development and safety testing of ion channels.
  • Conventional patch-clamp remains the preferred method for broader ion channel research.
  • Limitations exist in adapting robotic patch-clamp for diverse cell types and intracellular applications.

Purpose of the Study:

  • To demonstrate the expanded potential of robotic multiwell planar patch-clamp.
  • To establish protocols for efficient intracellular solution exchange in this system.
  • To assess the success rates of recordings from various native and primary mammalian cells.

Main Methods:

  • Cells were prepared as single-cell suspensions and robotically dispensed into microfluidic chambers with glass chips.
  • Automated control facilitated giga-seal formation, whole-cell access, and programmed voltage paradigms.
  • Fast extracellular and intracellular solution exchange protocols were implemented.

Main Results:

  • Successful recordings were achieved from diverse mammalian cell types, including those from blood vessels, joints, immune, and nervous systems.
  • The protocol demonstrated efficient intracellular solution exchange capabilities.
  • Recordings from 48 chambers yielded 16-33 cell recordings within 1-6 hours.

Conclusions:

  • Robotic multiwell planar patch-clamp is adaptable for a wider range of ion channel research applications.
  • The developed protocols enable efficient recordings from various primary and native cell types.
  • This technology offers a powerful tool for systematic ion channel analysis in diverse biological systems.