Inventors:
Ronald D. Paul - Fort Lauderdale FL, US
James L. Wyatt - San Diego CA, US
Barbara Carrillo - Miami FL, US
Oilda Rubio - Miami FL, US
Diana B. Careaga - Miami FL, US
Lidice L. Lopez - Miami FL, US
Assignee:
Beckman Coulter, Inc. - Fullerton CA
International Classification:
G01N 33/567
US Classification:
435 721, 435 723, 435 724, 4352852, 4352871, 4352872, 4352887, 436 10, 436 17, 436 56, 436164, 436171, 436175, 436546, 422 8201, 422 8205, 422 8207
Abstract:
A method useful for the enumeration of cell populations in a biological sample includes the steps of reacting in a single reaction mixture a sample, a first antibody labeled with a fluorochrome having a first emission spectrum and an additional antibody. The first antibody binds to an antigenic determinant differentially expressed on leukocytes and non-leukocytes. The additional antibody binds to an antigenic determinant differentially expressed on mature and immature granulocytes or myeloid cells, and is labeled either with the first fluorochrome or an additional fluorochrome having an emission spectrum distinguishable from the first emission spectrum. The reaction mixture can be mixed with a nucleic acid dye having an emission spectrum that overlaps with one of the first or additional emission spectra. The reaction mixture may be treated with a lytic system that differentially lyses non-nucleated red blood cells and conserves leukocytes. Populations of hematological cells are detected and enumerated using at least two parameters (fluorescence, optical, and electrical) for each population.