Method for measuring and evaluating the vitality of microorganisms and its applications

摘要

A method for assessing microorganism viability, including: (a) loading the microorganisms with a fluorescent substance, and (b) measuring the brightness of the intracellular fluorescence by flow cytometry. The method is characterised in that it includes the steps of: (i) incubating said microorganisms for 5-20 minutes with the marking substance in a concentration of 1-300 mu M and in the presence of a marking buffer at a pH of 4-8, to give microorganisms loaded with a fluorescent substance (loading step (a)); (ii) removing all of the marking substance not incorporated into the microorganisms; (iii) adding a source of energy prior to step (b); and (iv) using flow cytometry to measure the time required for half of the intracellular fluorescence of the microorganisms to disappear, within a predetermined brightness range, at a pH of 4-8 (outflow buffer) and at a temperature of 10-40 DEG C, preferably 20-40 DEG C.