Abstract

Fluorescent oligonucleotide hybridization probes were used to label bacterial cells for analysis by flow cytometry. The probes, complementary to short sequence elements within the 16S rRNA common to phylogenetically coherent assemblages of microorganisms, were labeled with tetramethylrhodamine and hybridized to suspensions of fixed cells. Flow cytometry was used to resolve individual target and nontarget bacteria (1 to 5 microns) via probe-conferred fluorescence. Target cells were quantified in an excess of nontarget cells. The intensity of fluorescence was increased additively by the combined use of two or three fluorescent probes complementary to different regions of the same 16S rRNA.

Keywords

Flow cytometry16S ribosomal RNAOligonucleotideOligomer restrictionBiologyFluorescenceRibosomal RNAFluorescent labellingMolecular biologyBacteriaFluorescence in situ hybridizationFluorescence microscopeMolecular probeCytometryDNAGeneticsGeneChromosome

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Publication Info

Year
1990
Type
article
Volume
56
Issue
6
Pages
1919-1925
Citations
4003
Access
Closed

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Rudolf Amann, Brian J. Binder, Robert Olson et al. (1990). Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations. Applied and Environmental Microbiology , 56 (6) , 1919-1925. https://doi.org/10.1128/aem.56.6.1919-1925.1990

Identifiers

DOI
10.1128/aem.56.6.1919-1925.1990