Flow cytometric analysis of Lactobacillus plantarum to monitor lag times, cell division and injury.
Flow cytometry in combination with fluorescent molecular markers 5- (and 6-) carboxyfluorescein succinimidylester (CFSE) and propidium iodide (PI) have been applied to determine lag times, numbers of cell divisions and injury after mild heat (50 degrees C, 5 min) and nisin treatments (0.1 and 1.0 microgram ml-1) of Lactobacillus plantarum. Initial labelling with covalently bound dye CFSE (20 and 100 micrograms ml-1) allowed determination of lag times and cell proliferation for up to eight generations. Double-labelling with CFSE and PI (5 micrograms ml-1) provided additional information about damage levels and distributions within populations. Subpopulations surviving treatment could be identified easily and selectively sorted.
Ueckert
J E
JE
Unilever Research Laboratorium, Vlaardingen, The Netherlands.
Nebe von-Caron
G
G
Bos
A P
AP
ter Steeg
P F
PF
D002455:Cell Division / Q000187:drug effects; D005434:Flow Cytometry; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005516:Food Microbiology; D005519:Food Preservation / Q000379:methods; D006358:Hot Temperature; D007700:Kinetics; D007778:Lactobacillus / Q000187:drug effects / Q000254:growth & development* / Q000302:isolation & purification; D009561:Nisin / Q000494:pharmacology; D011419:Propidium; D013388:Succinimides
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides; D009561:Nisin; D011419:Propidium