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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Quantifying Staphylococcus aureus Membrane Potential Using Flow Cytometry.

Neal D Hammer1

  • 1Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI, USA. hammern2@msu.edu.

Methods in Molecular Biology (Clifton, N.J.)
|July 15, 2021
PubMed
Summary
This summary is machine-generated.

This study quantifies membrane potential (MP) in Staphylococcus aureus using the cyanine dye DiOC and flow cytometry. This method helps understand bacterial physiology and antibiotic resistance mechanisms.

Keywords:
DiOCFlow cytometryMembrane potentialProton motive force (PMF)RespirationStaphylococcus aureus

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Area of Science:

  • Microbiology
  • Cell Biology
  • Biophysics

Background:

  • Membrane potential (MP) is crucial for bacterial physiology, impacting ATP generation, ion homeostasis, and antibiotic resistance.
  • Proton motive force (PMF) comprises MP and ΔpH, both vital for cellular functions.
  • Staphylococcus aureus relies on MP for critical processes, including energy production and survival.

Purpose of the Study:

  • To present a protocol for quantifying membrane potential (MP) in Staphylococcus aureus.
  • To utilize the cyanine dye diethyloxacarbocyanine iodide (DiOC) and flow cytometry for MP measurement.
  • To enable researchers to investigate factors affecting MP in S. aureus.

Main Methods:

  • Flow cytometry was employed to quantify fluorescent markers in cell populations.
  • The cyanine dye DiOC was used for its spectral and physical properties to assess MP.
  • A protocol was developed exploiting DiOC and flow cytometry to measure MP in S. aureus.

Main Results:

  • The study successfully quantified membrane potential (MP) in Staphylococcus aureus.
  • The developed assay allows for the measurement of MP using DiOC and flow cytometry.
  • This method provides a means to study the electron transport chain and factors influencing MP.

Conclusions:

  • The DiOC-based flow cytometry assay is an effective method for quantifying MP in S. aureus.
  • Understanding MP is essential for elucidating bacterial physiology and antibiotic resistance.
  • This protocol offers a valuable tool for future research on S. aureus and related bacteria.