title: Robotic multiwell planar patch-clamp for native and primary mammalian cells.
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.
authors.affiliation: Institute of Membrane and Systems Biology, Faculty of Biological Sciences, Garstang Building, University of Leeds, Leeds LS29JT, UK.
authors.forename: Carol J
authors.initials: CJ
authors.lastname: Milligan
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Piruthivi
authors.initials: P
authors.lastname: Sukumar
authors.forename: Yasser
authors.initials: Y
authors.lastname: Majeed
authors.forename: Mark L
authors.initials: ML
authors.lastname: Dallas
authors.forename: Anne
authors.initials: A
authors.lastname: English
authors.forename: Paul
authors.initials: P
authors.lastname: Emery
authors.forename: Karen E
authors.initials: KE
authors.lastname: Porter
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Smith
authors.forename: Ian
authors.initials: I
authors.lastname: McFadzean
authors.forename: Dayne
authors.initials: D
authors.lastname: Beccano-Kelly
authors.forename: Yahya
authors.initials: Y
authors.lastname: Bahnasi
authors.forename: Alex
authors.initials: A
authors.lastname: Cheong
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Naylor
authors.forename: Fanning
authors.initials: F
authors.lastname: Zeng
authors.forename: Xing
authors.initials: X
authors.lastname: Liu
authors.forename: Nikita
authors.initials: N
authors.lastname: Gamper
authors.forename: Lin-Hua
authors.initials: LH
authors.lastname: Jiang
authors.forename: Hugh A
authors.initials: HA
authors.lastname: Pearson
authors.forename: Chris
authors.initials: C
authors.lastname: Peers
authors.forename: Brian
authors.initials: B
authors.lastname: Robertson
authors.forename: David J
authors.initials: DJ
authors.lastname: Beech
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D056656:Lab-On-A-Chip Devices; D008214:Lymphocytes / Q000502:physiology; D032389:Myocytes, Smooth Muscle / Q000502:physiology; D018408:Patch-Clamp Techniques / Q000295:instrumentation* / Q000379:methods; D051381:Rats; D012371:Robotics / Q000295:instrumentation* / Q000379:methods