Patent ID: 7625712

Claim:
A method for the differentiation or enumeration of cell populations in a biological sample, said method comprising: A. passing through a flow aperture in a transducer that is capable of making multiple correlated measurements on cells, a mixture comprising: i. said biological sample; ii. a first antibody that binds to an antigenic determinant that is differentially expressed on leukocytes and non-leukocyte cells, said antibody labeled with a first fluorochrome having a first peak emission spectrum, iii. at least one additional antibody, which is (a) an additional antibody labeled with a fluorochrome having said first peak emission spectrum, wherein said additional antibody binds to an antigenic determinant that is differentially expressed on mature and immature granulocytes or myeloid cells; (b) an additional antibody labeled with an additional fluorochrome, wherein said additional antibody binds to an antigenic determinant that is differentially expressed on mature and immature granulocytes or myeloid cells, and wherein said additional fluorochrome has a peak emission spectrum distinguishable from said first peak emission spectrum; or (c) an additional antibody labeled with an additional fluochrome, wherein said additional antibody binds to an antigenic determinant that is differentially expressed on mature and immature granulocytes or myeloid cells, and wherein said additional fluorochrome has a peak emission spectrum that overlaps the first peak emission spectrum; and iv. a nucleic acid dye which has a peak emission spectrum that overlaps the peak emission spectrum of at least one of said first fluorochrome and said additional fluorochrome; B. detecting the fluorescence signals of said fluorochromes and nucleic acid dye and detecting at least one additional parameter which is an optical parameter, an electrical parameter, or combinations thereof, for the cells in the sample; and C. analyzing the fluorescence of said nucleic acid dye and said fluorochromes at an emission wavelength within the overlapping peak emission spectra that reveals additive fluorescence, with said at least one additional parameter to differentiate or enumerate populations of hematological cells in said biological sample.