Flow cytometry in environmental microbiology: A rapid approach for the isolation of single cells for advanced molecular biology analysis

Belinda C. Ferrari*, Tristrom J. Winsley, Peter L. Bergquist, Josie Van Dorst

*Corresponding author for this work

    Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

    4 Citations (Scopus)

    Abstract

    The isolation and subsequent characterization of microbial cells from within environmental samples is a difficult process. Flow cytometry and cell sorting, when combined with the application of fluorescent probes, have the capability for the detection and separation of diverse microbial populations from within complex mixtures. The isolation of single cells allows for downstream investigations towards system-level characterization of unknown Bacterial Phyla to occur. We describe here the combination of fluorescent in situ hybridization and cell sorting for the detection and isolation of Candidate Division TM7 bacteria from an enriched soil sample. The result is the isolation of rare cells suitable for advanced molecular analysis including whole genome amplification and high-throughput pyrosequencing.

    Original languageEnglish
    Title of host publicationMicrobial systems biology
    Subtitle of host publicationmethods and protocols
    EditorsAli Navid
    Place of PublicationNew York
    PublisherHumana Press Inc.
    Pages3-26
    Number of pages24
    Volume881
    ISBN (Electronic)9781617798276
    ISBN (Print)9781617798269
    DOIs
    Publication statusPublished - 2012

    Publication series

    NameMethods in Molecular Biology
    Volume881
    ISSN (Print)1064-3745

    Keywords

    • Cell sorting
    • Flow cytometry
    • Fluorescent in situ hybridization
    • Pyrosequencing
    • Single cell
    • Soil
    • Soil substrate membrane system
    • Whole genome amplification

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