title: Multiplex fluorescence in situ hybridization (M-FISH) and confocal laser scanning microscopy (CLSM) to analyze multispecies oral biofilms.
abstract: Multiplex fluorescence in situ hybridization (M-FISH) constitutes a favorable microbiological method for the analysis of spatial distribution of highly variable phenotypes found in multispecies oral biofilms. The combined use of confocal laser scanning microscopy (CLSM) produces high-resolution three-dimensional (3D) images of individual bacteria in their natural environment. Here, we describe the application of M-FISH on early (Streptococcus spp., Actinomyces naeslundii) and late colonizers (Fusobacterium nucleatum, Veillonella spp.) of in situ-formed oral biofilms, the acquisition of CLSM images, as well as the qualitative and quantitative analysis of these digitally obtained and processed images.
authors.affiliation: Department of Operative Dentistry and Periodontology, Albert-Ludwigs-University, Hugstetterstr, 55, 79106, Freidburg, Germany.
authors.forename: Lamprini
authors.initials: L
authors.lastname: Karygianni
authors.forename: Elmar
authors.initials: E
authors.lastname: Hellwig
authors.forename: Ali
authors.initials: A
authors.lastname: Al-Ahmad
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques; D018441:Biofilms; D002417:Cattle; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods; D009055:Mouth / Q000382:microbiology*