Open AccessUse of fluorescence ratio imaging microscopy and flow cytometry for estimation of cell vitality for Bacillus licheniformis
Author(s) -
Hornbæk Tina,
Dynesen Jens,
Jakobsen Mogens
Publication year - 2002
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2002.tb11400.x
Subject(s) - fluorescence microscope , flow cytometry , fluorescence , bacillus licheniformis , microscopy , fluorescence lifetime imaging microscopy , cytometry , intracellular , chemistry , biophysics , biology , microbiology and biotechnology , biochemistry , bacteria , pathology , optics , medicine , bacillus subtilis , physics , genetics
For Bacillus licheniformis SJ4628, an organism widely used in the enzyme industry, methods for determination of cell vitality at a single cell level using 5(6)‐carboxyfluorescein diacetate succinimidyl ester in combination with fluorescence ratio imaging microscopy and flow cytometry were developed. Immediately after inoculation and during growth, changes in intracellular pH values determined by fluorescence ratio imaging microscopy and in green fluorescence intensities determined by flow cytometry were observed. Correlations between the capacity to multiply and intracellular pH or green fluorescence intensity were demonstrated. Populations of cells not having a pH gradient or exhibiting low fluorescence intensities had significantly longer lag phases than populations of cells with a pH gradient and high fluorescence intensities.