Method for simultaneous analysis of cell viability, nucleated red blood cells and white blood cell differentials

摘要

A method for the simultaneous and quantitative, flow cytometric analysis of nucleated red blood cells (NRBC), white blood cells (WBC), damaged white blood cells and a white blood cell subclass differential (WBC/Diff) is provided. The method includes mixing an aliquot of a whole blood sample with a reagent system comprising a red blood cell (RBC) lysing component which lyses RBCs and NRBCs while minimizing damage to WBC cellular membranes and a membrane-impermeant nucleic acid stain which stains exposed NRBC nuclei and damaged WBCs, subjecting the stained aliquot to flow cytometric light measurements, obtaining at least one signal for the parameters of fluorescence (FL) and scattered light at a first and a second range of scatter angles, qualifying the obtained signals using AND/OR logic wherein to be qualified, a signal must be greater than the second scatter signal threshold AND also greater than either the first scatter signal threshold OR the FL threshold, constructing a three-dimensional plot of qualified intensity signals of fluorescence and scattered light from the detected signals, and differentiating the NRBC, WBC, damaged WBC and WBC/Diff from the constructed three-dimensional plot and determining the number of cells of each.