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Related Experiment Videos

A microflow superfusion system for use with excised membrane patches.

M Kakei, F M Ashcroft

    Pflugers Archiv : European Journal of Physiology
    |July 1, 1987
    PubMed
    Summary

    This study introduces a novel, rapid solution exchange method for excised membrane patches, requiring minimal solution volumes. This technique enables precise, fast solution changes for studying ion channel interactions with limited chemical availability.

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    Area of Science:

    • Biophysics
    • Electrophysiology
    • Membrane Biology

    Background:

    • Rapid solution exchange is crucial for studying ion channel kinetics.
    • Existing methods often require large solution volumes, limiting their application.
    • Investigating interactions with scarce compounds necessitates micro-volume techniques.

    Purpose of the Study:

    • To develop and validate a new method for rapid solution superfusion of excised membrane patches.
    • To enable high-throughput electrophysiology with minimal reagent consumption.
    • To facilitate the study of ion channel interactions using limited chemical quantities.

    Main Methods:

    • A novel perfusion system using a small tube and air bubbles for solution exchange.
    • Excised membrane patches superfused by inserting the pipette tip into the perfusion tube.

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  • Testing the method by measuring pipette resistance and K-channel currents during solution changes.

    • Main Results:

    • Solution exchange is a rapid step function, complete within 10-20 ms.
    • Recording interruption is minimal (<250 ms) and does not affect seal resistance.
    • The method effectively handles small solution volumes (approx. 100 µL).

    Conclusions:

    • This micro-volume rapid solution exchange method is efficient and reliable.
    • It is particularly advantageous for studying ion channel gating and pharmacology with limited reagents.
    • The technique enhances the study of single ionic channels and their interactions.