Journal of Industrial Microbiology and Biotechnology

, Volume 32, Issue 3, pp 94–102

New method to characterize microbial diversity using flow cytometry

Authors

  • Ho-Shin Park
    • Center for Environmental Science and Forensic ChemistryGas Technology Institute
    • Division of MicrobiologyKim Laboratories Inc.
  • Rebecca Schumacher
    • Center for Environmental Science and Forensic ChemistryGas Technology Institute
    • Center for Environmental Science and Forensic ChemistryGas Technology Institute
Original Paper

DOI: 10.1007/s10295-005-0208-3

Cite this article as:
Park, H., Schumacher, R. & Kilbane, J.J. J IND MICROBIOL BIOTECHNOL (2005) 32: 94. doi:10.1007/s10295-005-0208-3

Abstract

The majority of microorganisms have yet to be cultivated and represent a vast uncharacterized and untapped resource. Here, we report the utilization of a combination of flow cytometry, cultivation, and molecular genetics to develop new methodologies to access and characterize biodiversity in microbial samples. We demonstrate that fluorescent dyes and combinations of dyes can selectively stain portions of bacterial populations that can be isolated as sub-populations using fluorescence-activated cell sorting (FACS). Microbial sub-populations obtained by FACS differ substantially from the original microbial population, as demonstrated by denaturing gradient gel electrophoresis and determination of 16S rRNA gene sequences. These sub-populations can subsequently be used to inoculate microbial growth media, allowing the isolation of different microbial species from those that can be readily cultivated from the original sample using the same microbial growth media. When this technique was applied to the analysis of activated-sludge and Yellowstone Lake hydrothermal vent samples, comparative analysis of 16S rDNA sequences revealed that FACS allowed the detection of numerous bacterial species, including previously unknown species, not readily detectable in the original sample due to low relative abundance. This approach may result in a convenient methodology to more thoroughly characterize microbial biodiversity.

Keywords

Microbial diversityFlow cytometryUncultivatedCell sorting

Copyright information

© Society for Industrial Microbiology 2005