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In situ identification of streptococci and other bacteria in initial dental biofilm by confocal laser scanning microscopy and fluorescence in situ hybridization
Author(s) -
Dige Irene,
Nilsson Holger,
Kilian Mogens,
Nyvad Bente
Publication year - 2007
Publication title -
european journal of oral sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.802
H-Index - 93
eISSN - 1600-0722
pISSN - 0909-8836
DOI - 10.1111/j.1600-0722.2007.00494.x
Subject(s) - biofilm , in situ , fluorescence in situ hybridization , bacteria , microbiology and biotechnology , confocal , confocal laser scanning microscopy , dental plaque , in situ hybridization , biology , oligomer restriction , fluorescence microscope , 16s ribosomal rna , chemistry , fluorescence , oligonucleotide , biochemistry , optics , dna , genetics , physics , gene expression , organic chemistry , gene , chromosome
Confocal laser scanning microscopy (CLSM) has been employed as a method for studying intact natural biofilm. When combined with fluorescence in situ hybridization (FISH) it is possible to analyze spatial relationships and changes of specific members of microbial populations over time. The aim of this study was to perform a systematic description of the pattern of initial dental biofilm formation by applying 16S rRNA‐targeted oligonucleotide probes to the identification of streptococci and other bacteria, and to evaluate the usefulness of the combination of CLSM and FISH for structural studies of bacterial populations in dental biofilm. Biofilms were collected on standardized glass slabs mounted in intra‐oral appliances and worn by 10 individuals for 6, 12, 24 or 48 h. After intra‐oral exposure the biofilms were labelled with probes against either streptococci (STR405) or all bacteria (EUB338) and analysed by CLSM. The current approach of using FISH techniques enabled differentiation of streptococci from other bacteria and determination of their spatio‐temporal organization. The presence of chimney‐like multilayered microcolonies with different microbial compositions demonstrated by this methodology provided information supplementary to our previous knowledge obtained by classical electron microscopic methods and increased our understanding of the structure of developing biofilms.