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Measuring the Induced Membrane Voltage with Di-8-ANEPPS
05:52

Measuring the Induced Membrane Voltage with Di-8-ANEPPS

Published on: November 19, 2009

Measuring the induced membrane voltage with Di-8-ANEPPS.

Gorazd Pucihar1, Tadej Kotnik, Damijan Miklavcic

  • 1Department of Biomedical Engineering, Faculty of Electrical Engineering, University of Ljubljana. gorazd.pucihar@fe.uni-lj.si

Journal of Visualized Experiments : Jove
|November 21, 2009
PubMed
Summary

External electric fields induce a membrane voltage (DeltaPhi) in cells. This study demonstrates measuring DeltaPhi noninvasively using the fluorescent dye di-8-ANEPPS, showing cell shape influences voltage distribution.

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Area of Science:

  • Cellular Electrophysiology
  • Biophysics
  • Membrane Biology

Background:

  • External electric fields induce charge redistribution in cells, creating an induced membrane voltage (DeltaPhi).
  • This voltage superimposes on the resting membrane potential and exists only during field application.
  • Understanding DeltaPhi is crucial for cell electrophysiology and responses to electrical stimuli.

Purpose of the Study:

  • To present a protocol for noninvasively measuring the induced membrane voltage (DeltaPhi) in cells.
  • To demonstrate the utility of the potentiometric fluorescent dye di-8-ANEPPS for quantifying DeltaPhi.
  • To investigate the influence of cell shape on the amplitude and spatial distribution of DeltaPhi.

Main Methods:

  • Utilizing the potentiometric fluorescent dye di-8-ANEPPS to visualize and measure DeltaPhi.
  • Applying an external electric field to cells and observing fluorescence changes.
  • Analyzing fluorescence intensity changes, which are proportional to DeltaPhi variations.
  • Correlating cell morphology with the spatial distribution and magnitude of DeltaPhi.

Main Results:

  • Di-8-ANEPPS fluorescence intensity directly correlates with changes in DeltaPhi across the cell membrane.
  • The induced membrane voltage (DeltaPhi) can be measured noninvasively.
  • Cell shape significantly affects the amplitude and spatial distribution of the induced membrane voltage.
  • The protocol allows for real-time observation of DeltaPhi variations.

Conclusions:

  • Di-8-ANEPPS provides a reliable method for noninvasive measurement of induced membrane voltage (DeltaPhi).
  • Cellular electrophysiology is influenced by cell geometry, impacting responses to external electric fields.
  • This technique offers valuable insights into cell membrane potential dynamics under electrical stimulation.