Flow cytometry for better insight into the physiology of sporulating bacteria

Flow cytometry for better insight into the physiology of sporulating bacteria

Abstrakt

Conventional microbiological methods may not always be sufficient or appropriate when comes to an analysis of sporulating bacteria. A number of spores formed is usually evaluated via microscopic counting or by assessment of colony growth after heat treatment of cell culture. These methods are time-consuming and allow analysis of only a limited number of cells. Flow cytometry enables analysis of thousands of cells per second and thus enables rapid, simple and statistically relevant at-line analysis of a sporulating culture. Methodology for flow cytometry analysis of the spore-forming culture of the industrially interesting solventogenic bacterium Clostridium beijerinckiiwill be introduced. A population is distinguished into at least four subpopulations based on differences in light scatter and uptake or hydrolysis rate offluorescent dyes propidium iodide and carboxyfluorescein diacetate, reps., by various cell phenotypes. Together with the evaluation of physiological features, cells were subjected to extensive transcriptomic analysis. Examples, how flow cytometric data could be used for better characterization of individual stages of culture with differentially expressed genes will be presented.

Conventional microbiological methods may not always be sufficient or appropriate when comes to an analysis of sporulating bacteria. A number of spores formed is usually evaluated via microscopic counting or by assessment of colony growth after heat treatment of cell culture. These methods are time-consuming and allow analysis of only a limited number of cells. Flow cytometry enables analysis of thousands of cells per second and thus enables rapid, simple and statistically relevant at-line analysis of a sporulating culture. Methodology for flow cytometry analysis of the spore-forming culture of the industrially interesting solventogenic bacterium Clostridium beijerinckiiwill be introduced. A population is distinguished into at least four subpopulations based on differences in light scatter and uptake or hydrolysis rate offluorescent dyes propidium iodide and carboxyfluorescein diacetate, reps., by various cell phenotypes. Together with the evaluation of physiological features, cells were subjected to extensive transcriptomic analysis. Examples, how flow cytometric data could be used for better characterization of individual stages of culture with differentially expressed genes will be presented.

Clostridium beijerinckii; FC

Clostridium beijerinckii; FC

BRANSKÁ, B.; VASYLKIVSKA, M.; SEDLÁŘ, K.; JUREČKOVÁ, K.; PATÁKOVÁ, P.

08.09.2019

4th International Symposium on the Genetics of Industrial Microorganisms - Abstracts

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https://gim2019.org/fileadmin/user_upload/GIM2019/Abstractbook.pdf