Patent ID: 11970726
Assignee: DUTCH WATER TECHNOLOGIES B.V.
Field: Measurement (Instruments)
Classification: CPC C  G | IPC C  G

Claim 0:
1. A method for quantifying the cultivability of microbial cells using culture independent parameters, comprising the steps of:
(i) contacting a sample of microbial cells with a nucleic acid binding fluorescent dye to stain cellular nucleic acids in said microbial cells;
(ii) subjecting said microbial cells to a fluorescence microscopy imaging method;
(iii) determining at least a first parameter and a second parameter from the results of the fluorescence microscopy imaging method for each of said microbial cells, said first parameter being the nucleic acid content of said microbial cell and said second parameter being the extent of stretching of each of said microbial cells;
(iv) classifying, based on said at least first and second parameter, each of said microbial cells as either:, (A) a viable cultivable cell with a first parameter value with a fluorescence intensity between ≥0.1 and 1.0 and a second parameter value between ≥1.0 and 10 μm2,, (B) a viable non-cultivable cell with a first parameter value with a fluorescence intensity of ≥—0.2 and a second parameter value of ≥0.5 μm2, or, (C) a most likely dead cell with a first parameter value with a fluorescence intensity of <1.0 and a second parameter value of <0.5 μm2; and
(v) calculating a ratio for the sample, wherein the ratio is based on at least two of an amount of (A) viable cultivable cells in the sample, an amount of (B) viable non-cultivable cells in the sample and (C) an amount of most likely dead cells in the sample.