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An improved loose patch voltage clamp method using concentric pipettes.

W M Roberts, W Almers

    Pflugers Archiv : European Journal of Physiology
    |October 1, 1984
    PubMed
    Summary
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    This study introduces a novel noninvasive voltage-clamp technique for large cells, enabling accurate recordings even with low seal resistances. The method avoids enzymatic treatment and shows strong agreement with prior findings for sodium channels.

    Area of Science:

    • Cellular electrophysiology
    • Biophysical techniques

    Background:

    • Accurate voltage-clamp recordings are crucial for studying ion channel function.
    • Traditional methods often require enzymatic treatment or high seal resistances, limiting applications.

    Purpose of the Study:

    • To develop a noninvasive voltage-clamp method for large cells.
    • To achieve accurate and rapid recordings without enzymatic cell treatment.
    • To overcome limitations of low seal resistances in electrophysiological studies.

    Main Methods:

    • Utilized a dual-barrel, fire-polished pipette to isolate a cell membrane patch.
    • Electrically simulated a high-resistance seal by holding inner and outer regions isopotential.
    • Collected current from the inner region only for voltage-clamp analysis.

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  • Applied the technique to frog skeletal muscle and leech neurons.

    • Main Results:

    • Successfully performed noninvasive voltage-clamp recordings from large cells.
    • Achieved accurate and rapid measurements despite low seal resistances (approx. 1 MΩ).
    • Demonstrated good agreement with previously published data on frog muscle sodium channels.

    Conclusions:

    • The described method offers a valuable alternative for voltage-clamp recordings, particularly when enzymatic treatment is undesirable.
    • This technique expands the applicability of voltage-clamp studies to challenging cell types and conditions.
    • The findings validate the method's reliability and accuracy for ion channel research.