Robotic multiwell planar patch-clamp for native and primary mammalian cells.
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.
Milligan
Carol J
CJ
Institute of Membrane and Systems Biology, Faculty of Biological Sciences, Garstang Building, University of Leeds, Leeds LS29JT, UK.
Li
Jing
J
Sukumar
Piruthivi
P
Majeed
Yasser
Y
Dallas
Mark L
ML
English
Anne
A
Emery
Paul
P
Porter
Karen E
KE
Smith
Andrew M
AM
McFadzean
Ian
I
Beccano-Kelly
Dayne
D
Bahnasi
Yahya
Y
Cheong
Alex
A
Naylor
Jacqueline
J
Zeng
Fanning
F
Liu
Xing
X
Gamper
Nikita
N
Jiang
Lin-Hua
LH
Pearson
Hugh A
HA
Peers
Chris
C
Robertson
Brian
B
Beech
David J
DJ
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