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Perfusing pipettes.

J M Tang1, J Wang, F N Quandt

  • 1Department of Physiology, Rush Medical College, Chicago, IL 60612.

Pflugers Archiv : European Journal of Physiology
|May 1, 1990
PubMed
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A modified patch-clamp setup enables rapid fluid exchange in patch pipettes, completing in under one minute. This technique minimizes added noise, making it suitable for sensitive electrophysiological experiments.

Area of Science:

  • Electrophysiology
  • Cellular Biology
  • Biophysics

Background:

  • Patch-clamp electrophysiology is crucial for studying ion channel function.
  • Rapid solution exchange is often required for kinetic studies of channel gating and pharmacology.
  • Existing methods for solution exchange can be slow or introduce artifacts.

Purpose of the Study:

  • To develop and validate a simple modification to a patch-clamp setup for rapid fluid exchange.
  • To quantify the speed of solution exchange using a physiological indicator.
  • To assess the impact of the modification on system noise.

Main Methods:

  • A novel modification to the standard patch-clamp apparatus was implemented.
  • The rate of solution exchange was determined by measuring the reversal potential of calcium-activated potassium current.

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  • Noise analysis of the modified system was performed using power spectrum measurements.

    • Main Results:

    • The modified patch-clamp setup allows for complete solution exchange in less than 60 seconds.
    • The reversal potential changes rapidly, indicating efficient perfusion of the pipette interior.
    • The system introduces minimal excess noise below 1 kHz, preserving signal integrity.

    Conclusions:

    • This simple modification provides a fast and reliable method for solution exchange in patch-clamp recordings.
    • The technique is suitable for experiments requiring rapid application and removal of different solutions.
    • The low-noise characteristics ensure the quality of electrophysiological data.