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Force-controlled patch clamp of beating cardiac cells.
Journal article

Force-controlled patch clamp of beating cardiac cells.

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  • 2015-02-03
Published in:
  • Nano letters. - 2015
FluidFM
NaV1.5 channels
Whole-cell patch clamp
atomic force microscope
cardiomyocytes
microchanneled AFM cantilevers
Action Potentials
Equipment Design
Equipment Failure Analysis
Feedback
HEK293 Cells
Humans
Ion Channel Gating
Membrane Potentials
Micro-Electrical-Mechanical Systems
Micromanipulation
Microscopy, Atomic Force
Myocardial Contraction
Myocytes, Cardiac
NAV1.5 Voltage-Gated Sodium Channel
Patch-Clamp Techniques
Stress, Mechanical
English From its invention in the 1970s, the patch clamp technique is the gold standard in electrophysiology research and drug screening because it is the only tool enabling accurate investigation of voltage-gated ion channels, which are responsible for action potentials. Because of its key role in drug screening, innovation efforts are being made to reduce its complexity toward more automated systems. While some of these new approaches are being adopted in pharmaceutical companies, conventional patch-clamp remains unmatched in fundamental research due to its versatility. Here, we merged the patch clamp and atomic force microscope (AFM) techniques, thus equipping the patch-clamp with the sensitive AFM force control. This was possible using the FluidFM, a force-controlled nanopipette based on microchanneled AFM cantilevers. First, the compatibility of the system with patch-clamp electronics and its ability to record the activity of voltage-gated ion channels in whole-cell configuration was demonstrated with sodium (NaV1.5) channels. Second, we showed the feasibility of simultaneous recording of membrane current and force development during contraction of isolated cardiomyocytes. Force feedback allowed for a gentle and stable contact between AFM tip and cell membrane enabling serial patch clamping and injection without apparent cell damage.
Language
  • English
Open access status
closed
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Persistent URL
https://sonar.ch/global/documents/132005
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