PMID: 15227794Jul 2, 2004Paper

Obstacles to flow cytometric analysis of rumen microbial samples

Folia Microbiologica
L Lipoglavsek, G Avgustin

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.

References

May 1, 1988·Applied and Environmental Microbiology·D A StahlL Montgomery
Dec 1, 1972·The American Journal of Clinical Nutrition·M P Bryant
Apr 1, 1994·International Journal of Systematic Bacteriology·G AvgustinH J Flint
May 3, 2000·International Journal of Food Microbiology·B RegnaultP A Grimont
Sep 19, 2000·Journal of Microbiological Methods·A Moter, U B Göbel
May 26, 2001·Applied and Environmental Microbiology·K TajimaY Benno
Aug 15, 2001·Folia Microbiologica·L Lipoglavsek, G Avgustin
Mar 15, 2006·Applied and Environmental Microbiology·F Joux, P Lebaron

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