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Applications of flow cytometry in environmental microbiology and biotechnology

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Abstract

Flow cytometry (FCM) is a technique for counting, examining and sorting microscopic particles suspended in a stream of fluid. It uses the principles of light scattering, light excitation and the emission from fluorescent molecules to generate specific multiparameter data from particles and cells. The cells are hydrodynamically focussed in a sheath solution before being intercepted by a focused light source provided by a laser. FCM has been used primarily in medical applications but is being used increasingly for the examination of individual cells from environmental samples. It has found uses in the isolation of both culturable and hitherto non-culturable bacteria present infrequently in environmental samples using appropriate growth conditions. FCM lends itself to high-throughput applications in directed evolution for the analysis of single cells or cell populations carrying mutant genes. It is also suitable for encapsulation studies where individual bacteria are compartmentalised with substrate in water-in-oil-in-water emulsions or with individual genes in transcriptional/translational mixtures for the production of mutant enzymes. The sensitivity of the technique has allowed the examination of gene optimisation by a procedure known as random or neutral drift where screening and selection is based on the retention of some predetermined level of activity through multiple rounds of mutagenesis.

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Acknowledgments

Research at Macquarie was made possible by grants from the Australian Research Council, the Macquarie University Innovation Grants Fund and from Applimex Systems Pty Ltd.

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Authors and Affiliations

  1. Department of Chemistry and Biomolecular Sciences, Macquarie University, North Ryde, Sydney, NSW, 2109, Australia

    Peter L. Bergquist & Elizabeth M. Hardiman

  2. Department of Molecular Medicine & Pathology, University of Auckland Medical School, Auckland, New Zealand

    Peter L. Bergquist

  3. School of Biotechnology and Biomolecular Sciences, University of New South Wales, Randwick, NSW, 2052, Australia

    Belinda C. Ferrari & Tristrom Winsley

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  1. Peter L. Bergquist
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  2. Elizabeth M. Hardiman
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Correspondence to Peter L. Bergquist.

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Communicated by H. Santos.

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Bergquist, P.L., Hardiman, E.M., Ferrari, B.C. et al. Applications of flow cytometry in environmental microbiology and biotechnology. Extremophiles 13, 389–401 (2009). https://doi.org/10.1007/s00792-009-0236-4

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