Applications of flow cytometry in environmental microbiology and biotechnology

Peter L. Bergquist*, Elizabeth M. Hardiman, Belinda C. Ferrari, Tristrom Winsley

*Corresponding author for this work

    Research output: Contribution to journalReview articlepeer-review

    36 Citations (Scopus)

    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.

    Original languageEnglish
    Pages (from-to)389-401
    Number of pages13
    JournalExtremophiles
    Volume13
    Issue number3
    DOIs
    Publication statusPublished - May 2009

    Keywords

    • Fluorescence-activated cell sorting
    • Directed evolution
    • In vitro compartmentalisation
    • Non-culturable bacteria
    • Single cell analysis
    • Random (neutral) drift
    • Random mutagenesis
    • Protein optimisation
    • IN-VITRO COMPARTMENTALIZATION
    • WHOLE-CELL BIOSENSOR
    • DIRECTED EVOLUTION
    • MOLECULAR ANALYSIS
    • ENZYME EVOLUTION
    • DOUBLE EMULSIONS
    • QUANTUM DOTS
    • FLUORESCENCE
    • LIBRARIES
    • BACTERIA

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