Patent ID: 7332295

Claim:
A method for multiparameter identification, enumeration or characterization of cells in a sample, comprising: (a) obtaining a sample from an individual; (b) adding the sample to a container containing a pre-measured amount of counting particles; (c) adding to the sample a cocktail of fluorescently labeled monoclonal anti-HLADr, anti-CD33, anti-CD45 and anti-CD14 antibodies, wherein the fluorescent labels on the antibodies have peak emission spectra different from each other; (d) incubating the container to allow binding of the antibodies to the cells; (e) measuring fluorescence intensities and light scatter emissions of all cells bound to the fluorescently labeled monoclonal antibodies, and of the counting particles with an instrument capable of detecting and recording the intensities of at least four different fluorescence emissions, forward light scatter and sideward light scatter for each of the counting particles and for each of the bound cells in the sample; (f) storing measurement results as computer readable data for each counting particle and for each bound cell; (g) applying software algorithms which use logical combinations of gates to determine scattergram position and spread of total leukocytes, neutrophils, eosinophils, basophils, monocytes, lymphocytes, total dendritic cells, CD16-positive dendritic cells/monocytes, myeloid-derived dendritic cells and lymphoplasmacytoid dendritic cells, in addition to the counting particles, whereby each of the cell types is identified and quantitatively enumerated in terms of cells per unit of volume; and (f) applying software algorithms that use logical combinations of gates whereby clustering cell populations different from the normal leukocyte populations are identified.