title: Use of fluorescent staining and flow cytometry for monitoring physiological changes in solventogenic clostridia.
abstract: 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.
authors.affiliation: Department of Biotechnology, Institute of Chemical Technology in Prague, Technicka 5, 16628 Prague 6, Czech Republic. Electronic address: petra.patakova@vscht.cz.
authors.forename: Petra
authors.initials: P
authors.lastname: Patakova
authors.affiliation: Biopreparaty spol. s r. o., Na Vinicnich horach 37, 16000 Prague 6, Czech Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Linhova
authors.affiliation: Department of Biotechnology, Institute of Chemical Technology in Prague, Technicka 5, 16628 Prague 6, Czech Republic.
authors.forename: Pavla
authors.initials: P
authors.lastname: Vykydalova
authors.affiliation: Department of Biotechnology, Institute of Chemical Technology in Prague, Technicka 5, 16628 Prague 6, Czech Republic.
authors.forename: Barbora
authors.initials: B
authors.lastname: Branska
authors.affiliation: Department of Biotechnology, Institute of Chemical Technology in Prague, Technicka 5, 16628 Prague 6, Czech Republic.
authors.forename: Mojmir
authors.initials: M
authors.lastname: Rychtera
authors.affiliation: Department of Biotechnology, Institute of Chemical Technology in Prague, Technicka 5, 16628 Prague 6, Czech Republic.
authors.forename: Karel
authors.initials: K
authors.lastname: Melzoch
mesh_terms: D001463:Barbiturates; D061249:Batch Cell Culture Techniques; D019149:Bioreactors; D003013:Clostridium / Q000502:physiology* / Q000648:ultrastructure; D005285:Fermentation; D005434:Flow Cytometry; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007555:Isoxazoles; D011419:Propidium; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D001463:Barbiturates; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007555:Isoxazoles; C027780:carboxyfluoresceindiacetate; C052795:bis(1,3-dibutylbarbiturate)trimethine oxonol; D011419:Propidium
keywords: Clostridium beijerinckii; Clostridium tetanomorphum; Flow cytometry; Fluorescence staining