title: Obstacles to flow cytometric analysis of rumen microbial samples.
abstract: Several methods were tested that would improve the fluorescence signal from hybridized rumen bacterial cells. Disruption of cell envelopes by lysozyme, EDTA, proteinase K and/or SDS caused only a minor increase in fluorescence signal. Use of helper unlabeled oligonucleotide probes was successful only with the Puni[H672] probe which, however, when used with specific PBBl4-labeled probe, gave fluorescence signal drop. No substantial rise in fluorescence signal was also observed with cells subjected to growth-without-cell-division treatment. Further improvements are needed to make the fluorescent in situ hybridization (FISH)-flow cytometry combination applicable to rumen bacteria.
authors.affiliation: Zootechnical Department, Biotechnical Faculty, University of Ljubljana, 1230 Domzale, Slovenia.
authors.forename: L
authors.initials: L
authors.lastname: Lipoglavsek
authors.forename: G
authors.initials: G
authors.lastname: Avgustin
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D001431:Bacteriological Techniques; D015342:DNA Probes; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D017404:In Situ Hybridization, Fluorescence; D012417:Rumen / Q000382:microbiology*; D013194:Staining and Labeling
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes