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Multiparameter flow cytometry of bacteria.

Howard M Shapiro1, Gerhard Nebe-von-Caron

  • 1The Center for Microbial Cytometry and Howard M. Shapiro, MD, PC, West Newton, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 21, 2004
PubMed
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Flow cytometry can assess bacterial physiology by measuring membrane potential and permeability. These methods are reliable for both Gram-negative and Gram-positive bacteria, aiding in antimicrobial and environmental studies.

Area of Science:

  • Microbiology
  • Analytical Chemistry
  • Biotechnology

Background:

  • Bacterial size limits precision in flow cytometry measurements.
  • Differential interactions of bacteria with reagents complicate protocol design.
  • Standard multiparameter staining protocols are not universally applicable to diverse bacterial species.

Purpose of the Study:

  • To describe reliable flow cytometric methods for assessing bacterial physiologic states.
  • To differentiate methods for Gram-negative and Gram-positive bacteria.
  • To enable assessment of environmental and antimicrobial agent effects on microorganisms.

Main Methods:

  • Development of flow cytometric protocols tailored for bacterial analysis.
  • Measurement of membrane potential as an indicator of cell viability and function.

Related Experiment Videos

  • Measurement of membrane permeability to assess cell envelope integrity.

    • Main Results:

    • Established reliable flow cytometric techniques for Gram-negative bacteria.
    • Established reliable flow cytometric techniques for Gram-positive bacteria.
    • Demonstrated utility in evaluating effects of external factors on bacterial physiology.

    Conclusions:

    • Flow cytometry can reliably assess bacterial membrane potential and permeability.
    • Specific, validated methods are presented for Gram-negative and Gram-positive bacteria.
    • These techniques provide valuable insights into microbial responses to environmental changes and antimicrobial treatments.