Assessment of Gram- and Viability-Staining Methods for Quantifying Bacterial Community Dynamics Using Flow Cytometry

Frontiers in Microbiology
Aurore DuquenoyVincent Thomas

Abstract

Over the past years, gut microbiota became a major field of interest with increasing reports suggesting its association with a large number of human diseases. In this context, there is a major interest to develop analysis tools allowing simple and cost-effective population pattern analysis of these complex ecosystems to follow changes over time. Whereas sequence-based metagenomics profiling is widely used for microbial ecosystems characterization, it still requires time and specific expertise for analysis. Flow cytometry overcomes these disadvantages, providing key information on communities within hours. In addition, it can potentially be used to select, isolate and cultivate specific bacteria of interest. In this study, we evaluated the culturability of strictly anaerobic bacteria that were stained with a classical Live/Dead staining, and then sorted using flow cytometry under anaerobic conditions. This sorting of "viable" fraction demonstrated that 10-80% of identified "viable" cells of pure cultures of strictly anaerobic bacteria were culturable. In addition, we tested the use of a combination of labeled vancomycin and Wheat Germ Agglutinin (WGA) lectin to discriminate Gram-positive from Gram-negative bacteria in complex ec...Continue Reading

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Citations

Jan 21, 2021·MSystems·Peter Rubbens, Ruben Props

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Methods Mentioned

BETA
Flow Cytometry

Software Mentioned

vegan
FlowJo
Trimmomatic
phyloseq
R Software
24
FLASH
Micro
VSEARCH

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