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Related Experiment Videos

Membrane potential estimation by flow cytometry.

H M Shapiro1

  • 1283 Highland Avenue, West Newton, Massachusetts, 02465-2513, USA.

Methods (San Diego, Calif.)
|June 30, 2000
PubMed
Summary
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Membrane potential (delta psi) is crucial for cell function and integrity. Flow cytometry offers methods to measure this potential in various cell types, but careful technique is essential for accurate results.

Area of Science:

  • Cellular biology
  • Biophysics

Background:

  • Membrane potential (delta psi) is established by ion gradients and is vital for transmembrane signaling in many cell types.
  • Changes in delta psi are linked to cell development, function, pathology, and membrane integrity, particularly in bacteria lacking mitochondria.
  • Mitochondrial delta psi is a key indicator of cellular energy metabolism and is disrupted during apoptosis.

Purpose of the Study:

  • To review the role and measurement of membrane potential (delta psi) in various biological contexts.
  • To discuss the utility of flow cytometry for assessing delta psi in eukaryotic cells, mitochondria, and bacteria.
  • To highlight the limitations and necessary controls for accurate delta psi measurement.

Main Methods:

  • Utilizes flow cytometry for estimating membrane potential.

Related Experiment Videos

  • Employs lipophilic cationic and anionic dyes (e.g., cyanines, rhodamine 123, oxonols) for detecting changes in delta psi.
  • Incorporates newer ratiometric techniques for precise delta psi quantification.

    • Main Results:

    • Older dye-based methods detect large delta psi changes and population heterogeneity.
    • Newer ratiometric techniques enable precise delta psi measurement within 10 mV.
    • Identifies potential confounding factors affecting fluorescence measurements, such as efflux pumps and medium composition.

    Conclusions:

    • Accurate estimation and measurement of delta psi require careful control of experimental conditions.
    • Flow cytometry is a versatile tool for assessing membrane potential across different cell types and conditions.
    • Understanding delta psi is critical for interpreting cellular signaling, metabolism, and integrity.