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

Milligan, Carol J., Li, Jing, Sukumar, Piruthivi, Majeed, Yasser, Dallas, Mark L., English, Anne, Emery, Paul, Porter, Karen E., Smith, Andrew M., McFadzean, Ian, Beccano-Kelly, Dayne ORCID:, Bahnasi, Yahya, Cheong, Alex, Naylor, Jacqueline, Zeng, Fanning, Liu, Xing, Gamper, Nikita, Jiang, Lin-Hua, Pearson, Hugh A., Peers, Chris, Robertson, Brian and Beech, David J. 2009. Robotic multiwell planar patch-clamp for native and primary mammalian cells. Nature Protocols 4 , pp. 244-255. 10.1038/nprot.2008.230

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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.

Item Type: Article
Date Type: Published Online
Status: Published
Schools: Medicine
Publisher: Nature Research
ISSN: 1754-2189
Last Modified: 10 Nov 2022 11:20

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