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Flow Cytometry

Flow Cytometry

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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Related Experiment Video

Updated: Mar 14, 2026

An Optimized LIVE/DEAD Assay Coupled with Flow Cytometry for Quantifying Post-Stress Survival in Yeast Cells

An Optimized LIVE/DEAD Assay Coupled with Flow Cytometry for Quantifying Post-Stress Survival in Yeast Cells

Published on: August 29, 2025

Reevaluating multicolor flow cytometry to assess microbial viability.

Benjamin Buysschaert¹, Bo Byloos^1,2, Natalie Leys²

¹Centre for Microbial Ecology and Technology (CMET), Ghent University, Coupure Links 653, 9000, Ghent, Belgium.

Applied Microbiology and Biotechnology

|October 1, 2016

Summary

This summary is machine-generated.

Optimizing flow cytometry staining protocols is crucial for accurate bacterial viability assessment. This review discusses multicolor staining, dye optimization, and assay stability for reproducible results.

Keywords:

Functionality Multicolor FCM Protocols SYTO Viability

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Last Updated: Mar 14, 2026

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Molecular Probe Optimization to Determine Cell Mortality in a Photosynthetic Organism Microcystis aeruginosa Using Flow Cytometry

Molecular Probe Optimization to Determine Cell Mortality in a Photosynthetic Organism Microcystis aeruginosa Using Flow Cytometry

Published on: January 29, 2016

Area of Science:

Microbiology
Analytical Chemistry
Biotechnology

Background:

Flow cytometry offers rapid, quantitative bacterial viability analysis.
Current staining protocols are inconsistent, lacking general optimization strategies.
Multicolor flow cytometry protocols for microbiology are underdeveloped.

Purpose of the Study:

To review and discuss discrepancies in staining protocols for nucleic acid and functional dyes.
To present optimization strategies for multicolor flow cytometry in microbiology.
To highlight the importance of assay stability for reproducible results.

Main Methods:

Review of existing literature on flow cytometry staining protocols.
Discussion of cell-permeant nucleic acid and functional stains for viability.
Inclusion of original data on SYTO dyes (SYTO 59-64, SYTO 17) and functional stains.
Analysis of parameters influencing staining: kinetics, concentration, EDTA.

Main Results:

Discrepancies exist between protocols for different dye types.
SYTO dyes combined with functional stains enable double and triple staining.
Protocol tuning is essential due to dye-matrix-microorganism-instrument interactions.
EDTA can act as a membrane permeabilizer.

Conclusions:

Standardized, optimized multicolor flow cytometry protocols are needed for bacterial viability.
Further investigation into the stability of multicolor assays is critical.
Reproducible results require careful tuning of staining parameters for specific applications.