Flow cytometry in environmental microbiology: a rapid approach for the isolation of single cells for advanced molecular biology analysis.
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.
School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Randwick, NSW, Australia. b.ferrari@unsw.edu.au
Belinda C
BC
Ferrari
Tristrom J
TJ
Winsley
Peter L
PL
Bergquist
Josie
J
Van Dorst
D004783:Environmental Microbiology; D005434:Flow Cytometry / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence; D008967:Molecular Biology / Q000379:methods*