Protocol abstract


Nature Protocols 4, 244 - 255 (2009)
Published online: 5 February 2009 | doi:10.1038/nprot.2008.230

Subject Categories: Biochemistry and protein analysis | Pharmacology and toxicology

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

Carol J Milligan1, Jing Li1, Piruthivi Sukumar1, Yasser Majeed1, Mark L Dallas2, Anne English3, Paul Emery3, Karen E Porter2, Andrew M Smith4, Ian McFadzean5, Dayne Beccano-Kelly1, Yahya Bahnasi1,6, Alex Cheong1, Jacqueline Naylor1, Fanning Zeng1, Xing Liu1, Nikita Gamper1, Lin-Hua Jiang1, Hugh A Pearson1, Chris Peers2, Brian Robertson1 & David J Beech1


Robotic multiwell planar patch-clamp has become common in drug development and safety programs because it enables efficient and systematic testing of compounds against ion channels during voltage-clamp. It has not, however, been adopted significantly in other important areas of ion channel research, where conventional patch-clamp remains the favored method. Here, we show the wider potential of the multiwell approach with the ability for efficient intracellular solution exchange, describing protocols and success rates for recording from a range of native and primary mammalian cells derived from blood vessels, arthritic joints and the immune and central nervous systems. The protocol involves preparing a suspension of single cells to be dispensed robotically into 4–8 microfluidic chambers each containing a glass chip with a small aperture. Under automated control, giga-seals and whole-cell access are achieved followed by preprogrammed routines of voltage paradigms and fast extracellular or intracellular solution exchange. Recording from 48 chambers usually takes 1–6 h depending on the experimental design and yields 16–33 cell recordings.

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  1. Institute of Membrane and Systems Biology, Faculty of Biological Sciences, Garstang Building, University of Leeds, Leeds LS2 9JT, UK.
  2. Division of Cardiovascular and Neuronal Remodeling, Faculty of Medicine and Health, University of Leeds, Leeds LS2 9JT, UK.
  3. Academic Section of Musculoskeletal Disease, Leeds Institute of Molecular Medicine, School of Medicine, Chapel Allerton Hospital, University of Leeds, Chapeltown Road, Leeds LS7 4SA, UK.
  4. Department of Molecular Medicine, Rayne Institute, University College London, London, WC1E 6JJ, UK.
  5. King's College London, Division of Pharmaceutical Sciences Research, Sackler Institute of Pulmonary Pharmacology, School of Biomedical and Health Sciences, London SE1 1UL, UK.
  6. Department of Clinical Physiology, Faculty of Medicine, Minufiya University, Shibin El Kom, Monufia Governate, Egypt.

Correspondence to: David J Beech1 e-mail: d.j.beech@leeds.ac.uk

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