Increase in Fluorescence Intensity of 16S rRNA In Situ Hybridization in Natural Samples Treated with Chloramphenicol.

Applied and Environmental Microbiology
C C Ouverney, J A Fuhrman

Abstract

Despite the numerous advantages of fluorescent in situ hybridization for the identification of single prokaryotic cells with 16S rRNA probes, use of the technique with natural samples, especially those from the marine environment, is still problematic. The low percentage of fluorescently labeled cells constitutes the primary problem for in situ hybridization of natural samples, probably due to low cellular rRNA content. This study represents an attempt to improve detection of marine prokaryotes by increasing cellular rRNA content without changing the species composition. Cells from three California coastal sites were treated with chloramphenicol, an inhibitor of protein synthesis and rRNA degradation, at 100 (mu)g/ml and then were probed with a "universal" 16S rRNA fluorescent probe and viewed by image-intensified video microscopy. Counts of fluorescent cells increased from ca. 75% for untreated samples to ca. 93 to 99% for chloramphenicol-treated samples, compared to counts produced by DAPI (4(prm1),6-diamidino-2-phenylindole) staining, after at least 45 min of exposure to the drug (these percentages include autofluorescent cells, which averaged 6%). This suggests that most cells in these samples were active. We hypothesize th...Continue Reading

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Jun 1, 1995·Applied and Environmental Microbiology·U L Zweifel, A Hagstrom
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Citations

May 29, 2004·Extremophiles : Life Under Extreme Conditions·Antje Rusch, Jan P Amend
Dec 7, 2005·FEMS Microbiology Ecology·Guy C J Abell, John P Bowman
Feb 3, 2016·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·Ling ChenMary Rusckowski
Oct 31, 2000·Applied and Environmental Microbiology·C C Ouverney, J A Fuhrman
Mar 7, 2003·Applied and Environmental Microbiology·Mary M BrinigDavid A Relman
Jun 1, 2002·Applied and Environmental Microbiology·Annelie PernthalerRudolf Amann
Feb 10, 2004·Applied and Environmental Microbiology·Michinari SunamuraAkihiko Maruyama
Sep 9, 2005·Microbiology and Molecular Biology Reviews : MMBR·Jakob Pernthaler, Rudolf Amann
Oct 9, 2003·Applied and Environmental Microbiology·Cleber C OuverneyDavid A Relman
May 19, 2005·FEMS Microbiology Letters·Katrin Zwirglmaier
May 9, 1998·Applied and Environmental Microbiology·P LebaronP Catala

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