Simultaneous determination of bacterial viability and identity in biofilms using ethidium monoazide and fluorescent in situ hybridization
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 (...
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| Veröffentlicht in: | Water science and technology 2003-01, Vol.47 (5), p.123-128 |
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| container_issue | 5 |
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| container_title | Water science and technology |
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| creator | Regan, J M Oldenburg, P S Park, H D Harrington, G W Noguera, D R |
| description | 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. |
| doi_str_mv | 10.2166/wst.2003.0297 |
| format | Article |
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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. 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| fulltext | fulltext |
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| ispartof | Water science and technology, 2003-01, Vol.47 (5), p.123-128 |
| issn | 0273-1223 1996-9732 |
| language | eng |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals |
| subjects | Affinity Labels - chemistry Azides - chemistry Bacteria Biofilms Cell culture Dye dispersion Environmental Monitoring - methods ethidium monoazide Fish culture Fluorescence Fluorescence in situ hybridization Hybridization In Situ Hybridization, Fluorescence Membranes Microorganisms Nucleic acids Nucleic Acids - analysis Staining Target cells Viability |
| title | Simultaneous determination of bacterial viability and identity in biofilms using ethidium monoazide and fluorescent in situ hybridization |
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