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

Research output: Contribution to journalArticle

Carol J. Milligan, Jing Li, Piruthivi Sukumar, Yasser Majeed, Mark L. Dallas, Anne English, Paul Emery, Karen E. Porter, Andrew M. Smith, Ian McFadzean, Dayne Beccano-Kelly, Yahya Bahnasi, Alex Cheong, Jacqueline Naylor, Fanning Zeng, Xing Liu, Nikita Gamper, Lin-Hua Jiang, Hugh A. Pearson, Chris Peers & 2 more Brian Robertson, David J. Beech

Original languageEnglish
Pages (from-to)244 - 255
Number of pages12
JournalNature Protocols
Volume4
Issue number2
DOIs
Publication statusPublished - 2009

King's Authors

Abstract

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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