A combined immunofluorescence and fluorescent in situ hybridization assay for single cell analyses of dental plaque microorganisms.
A IF-FISH protocol was developed to characterize discrete cell populations within complex dental plaque samples. Optimizing and shortening the 3-step IF procedure and including RNase inhibitor in all IF reagents enabled the combination of both techniques and thus the simultaneous detection and enumeration of bacterial populations with distinct 16S rRNA sequences and/or surface markers of interest.
Gmür
Rudolf
R
Institute of Oral Biology, Section of Oral Microbiology and General Immunology, University of Zürich, Plattenstrasse 11, 8032 Zürich, Switzerland. gmuer@zzmk.unizh.ch
Lüthi-Schaller
Helga
H
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D003773:Dental Plaque / Q000382:microbiology*; D019085:Fluorescent Antibody Technique, Direct / Q000379:methods*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008856:Microscopy, Fluorescence; D010518:Periodontitis / Q000382:microbiology