Development of a rapid method for detecting bacterial cells in situ using 16S rRNA-targeted probes.
A rapid method for the identification of bacterial cells using 16S rRNA-directed, fluorescently tagged oligonucleotide probes has been developed. The parameters evaluated for their effect on labeling intensity included storage time, type of fixative, time of fixation, treatment time with methanol:formaldehyde and treatment time with borohydride. The results of tests using a variety of microorganisms, both Gram-positive and Gram-negative, are presented. Using this method, cells are spotted onto slides and stored desiccated until hybridized. This method may be especially applicable to environmental samples, which comprise diverse cell types and frequently require storage prior to examination.
D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification; D001431:Bacteriological Techniques; D005069:Evaluation Studies as Topic; D017403:In Situ Hybridization; D015336:Molecular Probe Techniques; D015345:Oligonucleotide Probes; D012329:RNA, Bacterial / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*
D015345:Oligonucleotide Probes; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S