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Analysis of Biofilm Matrix by Multiplex Fluorescence In Situ Hybridization (M-FISH) and Confocal Laser Scanning Microscopy (CLSM) During Nosocomial Infections

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Analytical Methodologies for Biofilm Research

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Abstract

Biofilm is the syntrophic consortium of the microorganisms that mainly comprises sessile communities of bacterial cells that adhere to inanimate or living surfaces with the help of extracellular polymeric substance (EPS) and pili by forming a matrix composed of exopolysaccharide, DNA, and proteins (polymeric conglomeration). The presence of various phenotypes within the biofilm can be determined with the help of the multiplex fluorescence in situ hybridization (M-FISH). High resolution 3D structures of the sessile communities found within the biofilm can be observed by the use of confocal laser scanning microscopy (CLSM). This chapter will focus on the application of M-FISH in studying the various types of sessile cells responsible for the development of the biofilm causing nosocomial diseases and the use of CLSM in the qualitative and quantitative analysis of the biofilm forming various nosocomial diseases.

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  1. Department of Biotechnology, Maulana Abul Kalam Azad University of Technology, West Bengal, Haringhata, India

    Bandita Dutta & Rina Rani Ray

  2. Department of Biotechnology, University of Engineering & Management, Kolkata, India

    Moupriya Nag & Dibyajit Lahiri

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  1. Department of Biotechnology, University of Engineering & Management, Kolkata, West Bengal, India

    Moupriya Nag

  2. Department of Biotechnology, University of Engineering & Management, Kolkata, West Bengal, India

    Dibyajit Lahiri

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Dutta, B., Nag, M., Lahiri, D., Ray, R.R. (2021). Analysis of Biofilm Matrix by Multiplex Fluorescence In Situ Hybridization (M-FISH) and Confocal Laser Scanning Microscopy (CLSM) During Nosocomial Infections. In: Nag, M., Lahiri, D. (eds) Analytical Methodologies for Biofilm Research. Springer Protocols Handbooks. Springer, New York, NY. https://doi.org/10.1007/978-1-0716-1378-8_8

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