Method for rapid and simultaneous analysis of nucleated red bloo

Chemistry: analytical and immunological testing – Biological cellular material tested

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436 10, 436 17, 436164, 436172, 436175, 435 2, 435 4, 435 29, 435 34, 435 39, 356 39, 356317, 356336, 356337, 356340, G01N 3348

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active

055590373

ABSTRACT:
A method and a device for the simultaneous and quantitative, flow cytometric analysis of nucleated red blood cells (NRBC) and white blood cells (WBC) in a whole blood sample. The method includes the lysis of red blood cells (RBC) and NRBC cytoplasm from an aliquot of a whole blood sample to eliminate RBC and to expose the NRBC to a vital nuclear stain and the inhibition of the permeation of the nuclear stain into the WBC, subjecting the stained aliquot to flow cytometric light measurements, obtaining at least one signal for parameters including scattered light at a first and a second range of scatter angles and fluorescence (FL), qualifying the obtained signals by using the combination logic wherein a qualified signal must be greater than the second scatter signal threshold, while at the same time it must be greater than either the first scatter signal threshold or the FL threshold {[(first scatter angle signal OR FL signals) AND second scatter angle signal]}, constructing a three-dimensional plot of qualified intensity signals of fluorescence and scattered light from the detected signals, and differentiating the NRBC and WBC from the constructed three-dimensional plot and determining the number of cells of each.

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