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Enumeration of bacteriophages using flow cytometry.

Corina P D Brussaard¹

¹Department of Biological Oceanography, Royal, Netherlands Institute for Sea Research, Texel, The Netherlands.

Methods in Molecular Biology (Clifton, N.J.)

|December 11, 2008

Summary

Flow cytometry now enables rapid bacteriophage enumeration, overcoming previous limitations in virus ecology studies. This method allows detailed discrimination of diverse viruses based on morphology, genome, and size.

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Area of Science:

Environmental microbiology
Virology
Molecular ecology

Background:

Bacteriophage enumeration was a significant challenge for virus ecology research.
Previous methods lacked the speed and sensitivity required for comprehensive studies.
Understanding bacteriophage populations is crucial for microbial ecosystem dynamics.

Purpose of the Study:

To present an optimized protocol for bacteriophage enumeration.
To leverage flow cytometry for rapid and sensitive virus detection.
To facilitate detailed characterization of bacteriophage communities.

Main Methods:

Utilized sensitive nucleic acid stains for virus detection.
Employed standard benchtop flow cytometry techniques.
Developed and optimized a protocol for bacteriophage enumeration.

Main Results:

The flow cytometric method allows detection and discrimination of various viruses.
The protocol enables enumeration of numerically important bacteriophages.
Different virus morphologies, genome types, and sizes can be distinguished.

Conclusions:

Flow cytometry offers a powerful tool for bacteriophage identification and enumeration.
This technique overcomes previous limitations in virus ecology studies.
The optimized protocol enhances the study of microbial ecosystem virus dynamics.