Simultaneous determination of bacterial viability and identity in biofilms using ethidium monoazide and fluorescent in situ hybridization

J. M. Regan, P. S. Oldenburg, H. D. Park, G. W. Harrington, D. R. Noguera

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

A protocol for simultaneously interrogating bacterial viability and identity using in situ, culture-independent methods is described. Viability is assayed using ethidium monoazide (EMA) staining of cells with compromised membranes, and identity is determined using fluorescent in situ hybridization (FISH). Experiments with planktonic cultures were used to demonstrate the compatibility of EMA staining and FISH after covalently bonding EMA to nucleic acids by photoreaction. Applications to biofilm samples showed that diffusion limitations in the biofilm matrix were not problematic and that effective discrimination of viable target cells within a mixed microbial community was possible.

Original languageEnglish
Pages (from-to)123-128
Number of pages6
JournalWater Science and Technology
Volume47
Issue number5
DOIs
Publication statusPublished - 2003
Externally publishedYes

Keywords

  • Biofilm
  • Ethidium monoazide
  • Fluorescent in situ hybridization
  • Viability

ASJC Scopus subject areas

  • Environmental Engineering
  • Water Science and Technology

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