A novel way to go whole-cell in patch-clamp experiments.
With a conventional patch-clamp electrode, an Ag/AgCl wire sits stationary inside the pipette. To move from the gigaseal cell-attached configuration to whole-cell recording, suction is applied inside the pipette. We have designed and developed a novel Pushpen patch-clamp electrode, in which a W wire insulated and wound with Ag/AgCl wire can move linearly inside the pipette. The W wire has a conical tip, which can protrude from the pipette tip like a push pen, a procedure we call the Pushpen Operation. We use the Pushpen operation to impale the cell membrane in cell-attached configuration to go whole-cell without disruption of the gigaseal. We successfully recorded whole-cell currents from chinese hamster ovarian cells expressing influenza A virus protein A/M2, after obtaining whole-cell configuration with the Pushpen operation. This novel method of achieving whole-cell configuration may have a higher success rate than is the case with the conventional patch clamp technique.
Inayat
Samsoon
S
Zhao
Yan
Y
Cantrell
Donal R
DR
Dikin
Dmitryi
D
Pinto
Lawrence H
LH
Troy
John B
JB
D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D003412:Cricetulus; D004594:Electrophysiology / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D006801:Humans; D018408:Patch-Clamp Techniques / Q000295:instrumentation* / Q000379:methods; D014763:Viral Matrix Proteins / Q000378:metabolism; D000086082:Viroporin Proteins
D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins