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Measurements of membrane patch capacitance using a software-based lock-in system.

Andreas Neef1, Christian Heinemann, Tobias Moser

  • 1InnerEarLab, Department of Otolaryngology, Goettingen University Medical School, 37099 Goettingen, Germany. aneef@gwdg.de

Pflugers Archiv : European Journal of Physiology
|January 9, 2007
PubMed
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This study introduces a new software lock-in technique for on-cell patch-clamp capacitance measurements, improving temporal resolution for studying vesicle fusion and fission dynamics in cells.

Area of Science:

  • Cell Biology
  • Biophysics
  • Electrophysiology

Background:

  • On-cell patch-clamp capacitance measurements track vesicle fusion events.
  • Traditional methods using hardware lock-in amplifiers have limited temporal resolution and underutilize patch current data.

Purpose of the Study:

  • To present a versatile, high-temporal-resolution approach for on-cell patch-clamp capacitance measurements.
  • To leverage digitally controlled patch-clamp amplifiers and software lock-in systems for enhanced data acquisition.

Main Methods:

  • Utilized a digitally controlled patch-clamp amplifier with a software lock-in system.
  • Recorded patch current alongside capacitance and conductance.
  • Performed high-resolution measurements on chromaffin cell patches.

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Main Results:

  • The software lock-in system demonstrated superior bandwidth and signal-to-noise ratio compared to hardware methods.
  • Vesicle fission was observed to complete within tens of microseconds.
  • Direct current analysis revealed subtle current changes during vesicle fusion and fission.

Conclusions:

  • The software lock-in approach offers a more versatile and higher temporal resolution method for patch capacitance measurements.
  • This technique provides novel insights into the membrane properties of fusing and fissions vesicles.
  • The study includes a practical guide for implementing these high-resolution measurements.