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

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 others Brian Robertson, David J. Beech

Research output: Contribution to journalArticle

66 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)244-255
Number of pages12
JournalNature Protocols
Volume4
Issue number2
DOIs
Publication statusPublished - 16 Feb 2009
Externally publishedYes

Fingerprint

Clamping devices
Robotics
Cells
Ion Channels
Ion exchange
Blood vessels
Neurology
Electric potential
Microfluidics
Design of experiments
Seals
Suspensions
Arthritis
Glass
Blood Vessels
Research Design
Central Nervous System
Joints
Testing
Safety

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Milligan, C. J., Li, J., Sukumar, P., Majeed, Y., Dallas, M. L., English, A., ... Beech, D. J. (2009). Robotic multiwell planar patch-clamp for native and primary mammalian cells. Nature Protocols, 4(2), 244-255. https://doi.org/10.1038/nprot.2008.230

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; Bahnasi, Yahya; Cheong, Alex; Naylor, Jacqueline; Zeng, Fanning; Liu, Xing; Gamper, Nikita; Jiang, Lin Hua; Pearson, Hugh A.; Peers, Chris; Robertson, Brian; Beech, David J.

In: Nature Protocols, Vol. 4, No. 2, 16.02.2009, p. 244-255.

Research output: Contribution to journalArticle

Milligan, CJ, Li, J, Sukumar, P, Majeed, Y, Dallas, ML, English, A, Emery, P, Porter, KE, Smith, AM, McFadzean, I, Beccano-Kelly, D, Bahnasi, Y, Cheong, A, Naylor, J, Zeng, F, Liu, X, Gamper, N, Jiang, LH, Pearson, HA, Peers, C, Robertson, B & Beech, DJ 2009, 'Robotic multiwell planar patch-clamp for native and primary mammalian cells', Nature Protocols, vol. 4, no. 2, pp. 244-255. https://doi.org/10.1038/nprot.2008.230
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 ; Bahnasi, Yahya ; Cheong, Alex ; Naylor, Jacqueline ; Zeng, Fanning ; Liu, Xing ; Gamper, Nikita ; Jiang, Lin Hua ; Pearson, Hugh A. ; Peers, Chris ; Robertson, Brian ; Beech, David J. / Robotic multiwell planar patch-clamp for native and primary mammalian cells. In: Nature Protocols. 2009 ; Vol. 4, No. 2. pp. 244-255.
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