Quantification of lipids and protein in thin biofilms by fluorescence staining

Biofouling
Poul LarsenJeppe Lund Nielsen

Abstract

The efficiency of removing unwanted biofilm from surfaces in industrial water systems was examined by fluorescence microscopy and image analysis. A quantitative assay for in situ determination of biofilm components was developed and tested on thin biofilms grown in reactors as well as real biofilms sampled from a fish processing factory. Different fluorescent dyes for in situ detection of protein, lipid and total organic matter were tested. It was possible to determine the approximate amounts, concentrations and coverage of the different components by correlating the fluorescent intensity of the biofilm components to standard solutions immobilised as a biofilm. The quantification methods were evaluated as a strategy for determining the efficiency of different disinfection/cleaning procedures, showing that quantification of these biofilm components was fast and reliable for optimisation of cleaning in place procedures. However, the approach also showed that bacterial cells, as investigated by culture-independent procedures, were killed but not removed by most disinfection procedures tested, potentially leading to surfaces which are easily recolonised.

References

Mar 1, 1985·The Journal of Cell Biology·P GreenspanS D Fowler
Aug 1, 1985·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·S D Fowler, P Greenspan
Jul 2, 1999·Applied and Environmental Microbiology·M V ZubkovR Amann
May 24, 2003·Journal of Industrial Microbiology & Biotechnology·Johanna MaukonenMaria Saarela
Jul 16, 2005·Biofouling·B V KjellerupP H Nielsen
Jun 20, 2006·Current Biology : CB·John M Ratcliffe, M Brock Fenton
Nov 10, 2007·Environmental Microbiology·Poul LarsenPer Halkjaer Nielsen

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Citations

May 5, 2011·Biotechnology Advances·R Lencastre FernandesK V Gernaey
Jan 17, 2012·FEMS Microbiology Ecology·Anja KristiansenJeppe L Nielsen
Jun 21, 2016·Biotechnology & Genetic Engineering Reviews·Neelam MangwaniSurajit Das
Dec 30, 2015·Applied and Environmental Microbiology·Philippe VogeleerJosée Harel
May 7, 2020·Methods and Applications in Fluorescence·Olena MoshynetsVladyslava Kovalska

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Methods Mentioned

BETA
Fluorescence
scraping
protein assay

Software Mentioned

NanoOrange
MetaVye
ImageJ

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