A steam-based method to investigate biofilm

Scientific Reports
Jason TasseFrédéric Laurent

Abstract

Biofilm has become a major topic of interest in medical, food, industrial, and environmental bacteriology. To be relevant, investigation of biofilm behavior requires effective and reliable techniques. We present herein a simple and robust method, adapted from the microplate technique, in which steam is used as a soft washing method to preserve biofilm integrity and to improve reproducibility of biofilm quantification. The kinetics of steam washing indicated that the method is adapted to remove both planktonic bacteria and excess crystal violet (CV) staining for S. aureus, S. epidermidis, S. carnosus, P. aeruginosa, and E. coli biofilm. Confocal laser scanning microscopy confirmed that steam washing preserved the integrity of the biofilm better than pipette-based washing. We also investigated the measurement of the turbidity of biofilm resuspended in phosphate-buffered saline (PBS) as an alternative to staining with CV. This approach allows the discrimination of biofilm producer strains from non-biofilm producer strains in a way similar to CV staining, and subsequently permits quantification of viable bacteria present in biofilm by culture enumeration from the same well. Biofilm quantification using steam washing and PBS turbidi...Continue Reading

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Citations

Sep 3, 2020·International Journal of Molecular Sciences·Alan DiotFrédéric Laurent
Nov 27, 2020·Frontiers in Medicine·Rossella Baldan, Parham Sendi
Aug 28, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Sylwia ZielińskaAdam Matkowski

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Methods Mentioned

BETA
Laser
ELISA

Software Mentioned

GraphPad
Prism
Imaris
Comstat2

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