Title :
A Novel Way to Go Whole Cell in Patch-Clamp Experiments
Author :
Inayat, Samsoon ; Zhao, Yan ; Cantrell, Donald R. ; Dikin, Dmitriy A. ; Pinto, Lawrence H. ; Troy, John B.
Author_Institution :
Dept. of Biomed. Eng., Northwestern Univ., Evanston, IL, USA
Abstract :
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 the conventional patch-clamp technique.
Keywords :
bioelectric phenomena; biological specimen preparation; biomembranes; cellular biophysics; electrodes; proteins; Ag/AgCl wire; Chinese hamster ovarian cells; Pushpen operation; Pushpen patch-clamp electrode; W wire; cell membrane; gigaseal cell-attached configuration; influenza A virus protein A/M2; patch-clamp experiments; pipette; suction; whole cell; Electrophysiology; gigaseal; patch clamp; whole cell;
Journal_Title :
Biomedical Engineering, IEEE Transactions on
DOI :
10.1109/TBME.2010.2055055