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

3 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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