title: Force-controlled patch clamp of beating cardiac cells.
abstract: 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.
authors: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich , Zurich, Switzerland.
authors: Dario
authors: D
authors: Ossola
authors: Mohamed-Yassine
authors: MY
authors: Amarouch
authors: Pascal
authors: P
authors: Behr
authors: János
authors: J
authors: Vörös
authors: Hugues
authors: H
authors: Abriel
authors: Tomaso
authors: T
authors: Zambelli
mesh_terms: D000200:Action Potentials / Q000502:physiology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005246:Feedback; D057809:HEK293 Cells; D006801:Humans; D015640:Ion Channel Gating / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology*; D055617:Micro-Electrical-Mechanical Systems / Q000295:instrumentation*; D008846:Micromanipulation / Q000295:instrumentation; D018625:Microscopy, Atomic Force / Q000295:instrumentation; D009200:Myocardial Contraction / Q000502:physiology; D032383:Myocytes, Cardiac / Q000502:physiology*; D062554:NAV1.5 Voltage-Gated Sodium Channel / Q000502:physiology*; D018408:Patch-Clamp Techniques / Q000295:instrumentation*; D013314:Stress, Mechanical
chemical_list: D062554:NAV1.5 Voltage-Gated Sodium Channel; C568320:SCN5A protein, human
keywords: FluidFM; NaV1.5 channels; Whole-cell patch clamp; atomic force microscope; cardiomyocytes; microchanneled AFM cantilevers