Use of fluorescent staining and flow cytometry for monitoring physiological changes in solventogenic clostridia
Language English Country England, Great Britain Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
24211310
DOI
10.1016/j.anaerobe.2013.10.006
PII: S1075-9964(13)00173-X
Knihovny.cz E-resources
- Keywords
- Clostridium beijerinckii, Clostridium tetanomorphum, Flow cytometry, Fluorescence staining,
- MeSH
- Barbiturates MeSH
- Staining and Labeling methods MeSH
- Bioreactors MeSH
- Clostridium physiology ultrastructure MeSH
- Fermentation * MeSH
- Fluoresceins MeSH
- Fluorescent Dyes MeSH
- Isoxazoles MeSH
- Propidium MeSH
- Flow Cytometry MeSH
- Batch Cell Culture Techniques MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Barbiturates MeSH
- bis(1,3-dibutylbarbiturate)trimethine oxonol MeSH Browser
- carboxyfluoresceindiacetate MeSH Browser
- Fluoresceins MeSH
- Fluorescent Dyes MeSH
- Isoxazoles MeSH
- Propidium MeSH
Physiological changes in populations of Clostridium beijerinckii and Clostridium tetanomorphum were monitored by fluorescence staining and flow cytometry. To estimate the number of metabolically active cells in exponential growth, a combination of the dyes propidium iodide and carboxy fluorescein diacetate appeared to be a good choice for both species. During stationary phase, these stains did not reflect physiological changes sufficiently and therefore additional labeling with bis-(1,3-dibutylbarbituric acid) trimethineoxonol was applied. Results of fluorescence staining in solventogenic batch fermentations were compared with substrate-use data, the concentration of key metabolites and growth curves. We demonstrate that measurements by all methods were mutually compatible.
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