• DocumentCode
    1525746
  • 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
  • Volume
    57
  • Issue
    11
  • fYear
    2010
  • Firstpage
    2764
  • Lastpage
    2770
  • 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;
  • fLanguage
    English
  • Journal_Title
    Biomedical Engineering, IEEE Transactions on
  • Publisher
    ieee
  • ISSN
    0018-9294
  • Type

    jour

  • DOI
    10.1109/TBME.2010.2055055
  • Filename
    5497104