Population patch clamp improves data consistency and success rates in the measurement of ionic currents.
Present whole-cell patch-clamp methodology has only moderate consistency and throughput, rendering impractical functional measurements on large numbers of ion channel ligands or on large numbers of unknown or mutant channel genes. In the population patch clamp (PPC) described herein, a single voltage-clamp amplifier sums the whole-cell currents from multiple cells at once, each sealed to a separate aperture in a planar substrate well. The resulting ensemble currents are more consistent from well to well, and the success rate for each recording attempt is >95%. The PPC was implemented by modifying the PatchPlate substrate and amplifiers in the IonWorks patch-clamp instrument. The increased data consistency and likelihood of a successful recording in each well, combined with 384-well measurements in parallel, allow the direct electrophysiological recording of thousands of ensemble ionic currents per day. Therapeutic groups in drug discovery programs require this order of throughput to screen directed compound libraries against ion channel targets. The potential for studying the function of large numbers of ion channel mutants may be realized with the technique. The procedure incorporates subtraction methods that correct for expected distortions and also reliably produces data that agree with previous patch-clamp studies.
Finkel
Alan
A
Molecular Devices Corporation, Union City, California 94587, USA.
Wittel
Andrew
A
Yang
Naibo
N
Handran
Shawn
S
Hughes
Jan
J
Costantin
James
J
D015761:4-Aminopyridine / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D002463:Cell Membrane Permeability / Q000187:drug effects; D003198:Computer Simulation; D006224:Cricetinae; D000072237:ERG1 Potassium Channel; D004594:Electrophysiology / Q000379:methods*; D051638:Ether-A-Go-Go Potassium Channels / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D051664:Kv1.3 Potassium Channel / Q000187:drug effects / Q000378:metabolism; D008012:Lidocaine / Q000494:pharmacology; D008954:Models, Biological; D009124:Muscle Proteins / Q000187:drug effects / Q000378:metabolism; D062554:NAV1.5 Voltage-Gated Sodium Channel; D018408:Patch-Clamp Techniques / Q000379:methods* / Q000592:standards*; D024683:Potassium Channels, Tandem Pore Domain / Q000187:drug effects / Q000378:metabolism; D015203:Reproducibility of Results; D015222:Sodium Channels / Q000187:drug effects / Q000378:metabolism; D013748:Tetracaine / Q000494:pharmacology
D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; C097044:KCNAB1 protein, human; D051664:Kv1.3 Potassium Channel; D009124:Muscle Proteins; D062554:NAV1.5 Voltage-Gated Sodium Channel; D024683:Potassium Channels, Tandem Pore Domain; C568320:SCN5A protein, human; D015222:Sodium Channels; D013748:Tetracaine; D008012:Lidocaine; D015761:4-Aminopyridine