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Multiplex Fluorescence In Situ Hybridization (M-FISH) and Confocal Laser Scanning Microscopy (CLSM) to Analyze Multispecies Oral Biofilms

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Microbial Biofilms

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1147))

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.

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Correspondence to Ali Al-Ahmad .

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Karygianni, L., Hellwig, E., Al-Ahmad, A. (2014). Multiplex Fluorescence In Situ Hybridization (M-FISH) and Confocal Laser Scanning Microscopy (CLSM) to Analyze Multispecies Oral Biofilms. In: Donelli, G. (eds) Microbial Biofilms. Methods in Molecular Biology, vol 1147. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0467-9_5

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  • DOI: https://doi.org/10.1007/978-1-4939-0467-9_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0466-2

  • Online ISBN: 978-1-4939-0467-9

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