Patent ID: 8563251

Claim:
A method for remote enumerating of microbial cells in a sample using a multiplexed high through-put system designed for remote measuring of total fluorescence of the sample to estimate the number of cells in the sample, the method comprising the following steps: (a) providing a multiplexed high through-put system designed for remote measuring of total fluorescence of the sample, a fluorescent cyanine nucleic acid stain, a DNase I, and a liquid or a water sample, wherein the multiplexed high through-put system comprises: at least one sample holding unit, mixing unit and injection unit, at least two low protein binding filters for filtering the samples, wherein the system comprises at least one 0.45 μm low protein binding filter for samples and at least one 0.02 μm low protein binding filter for blanks, an injection unit for injecting into the mixing unit the cyanine nucleic acid stain and DNase I, wherein the mixing unit mixes the cyanine nucleic acid stain and DNase I, or the injection unit injects and mixes the cyanine nucleic acid stain and DNase I, a sample processor for inputting samples into the fluorometer, a fluorometer, a data recorder, a data output device for outputting to a user a total fluorescence, and a computer system for calculating a total fluorescence; (b) filtering the liquid or water sample, and a liquid or water blank, with at least one low protein binding filter, wherein the 0.45 μm filter filters samples and the 0.02 μm filter filters blanks; (c) adding the DNase I and the fluorescent cyanine nucleic acid stain, or equivalent, to the filtered sample and filtered blank, wherein the mixing unit mixes the cyanine nucleic acid stain and DNase I, or the injection unit injects and mixes the cyanine nucleic acid stain and DNase I; (d) measuring a total bulk fluorescence in the sample and the blank, wherein the total fluorescence is measured using an excitation wavelength of 495 lambda and an emission spectra ranging from 450 to 650 lambda; (e) outputting to a user by the data output device the measured total fluorescence; and (f) calculating using the computer system the total fluorescence by comparing the measured total fluorescence in the 0.45 μm filtered cell-comprising sample with the 0.02 μm filtered blank control sample, and determining the estimate of the total number of microbial cells in the sample by correlating the total fluorescence to a standard sample measurement.