Abstract
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 stan-
dardized 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 compo-
sitions 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.
| Original language | English |
|---|---|
| Journal | European Journal of Oral Sciences |
| Volume | 115 |
| Pages (from-to) | 459-467 |
| Number of pages | 9 |
| ISSN | 0909-8836 |
| Publication status | Published - 2007 |
Keywords
- biofilm
- confocal laser scanning
- dental plaque;
- fluorescence in situ
- ultrastructure