In situ identification of streptococci and other bacteria in initial dental biofilm by confocal laser scanning microscopy and fluorescence in situ hybridization

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  • Danish Biomembrane Research Centre
  • Department of Dental Pathology, Operative Dentistry and Endodontics
  • Department of Medical Microbiology and Immunology

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 languageEnglish
JournalEuropean Journal of Oral Sciences
Pages (from-to)459-467
Number of pages9
Publication statusPublished - 2007

    Research areas

  • biofilm, confocal laser scanning, dental plaque;, fluorescence in situ, ultrastructure

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