Beckman Coulter, Inc.

Ravindra Mylvaganam

Ronald D Paul

Diana B Careaga

Lidice L Lopez

Oilda Rubio

A method for identifying a platelet population, preferably a population of immature, reticulated platelets, in a biological sample involves incubating the biological sample for less than 5 minutes with at least one labeled, ligand (e.g., monoclonal antibody) that binds to an epitope or antigen on platelets and with a nucleic acid dye. In one embodiment, the dye is Acridine Orange and the label on the ligand is PE-Cy7. The sample is then analyzed and one or more platelet populations is rapidly identified or quantified by passing the incubated sample through a sensing region of a flow cytometer. In one embodiment, this method occurs without a washing or physical cell separation step. The incubated sample is irradiated with a laser light source, and fluorescence of the labeled ligand and the nucleic acid dye are measured along with at least one additional parameter, e.g., light scatter, direct current, axial light loss, opacity, radio frequency, and fluorescence. These parameters are used to identify qualitatively or quantitatively the platelet populations in the sample. This rapid analytic method is particularly valuable in clinical situations where either low platelet counts or interfering conditions lead to inaccuracies of the platelet measurement. This method is suitable for performance in an automated hematology analyzer.

Method for a rapid antibody-based analysis of platelet populations

Methods for differentially identifying cells in an instrument employ compositions containing a combination of selected antibodies and fluorescent dyes having different cellular distribution patterns and specificities, as well as antibodies and fluorescent dyes characterized by overlapping emission spectra which form non-compensatable spectral patterns. When utilizing the compositions described herein consisting of fluorescent dyes and fluorochrome labeled antibodies with overlapping spectra that cannot be separated or distinguished based upon optical or electronic compensation means, a new fluorescent footprint is established. This new fluorescent footprint is a result of the overlapping spectra and the combined cellular staining patterns of the dyes and fluorochrome labeled antibodies chosen for the composition. The new fluorescent footprint results in histogram patterns that are useful for the identification of additional cell populations or subtypes in hematological disease.

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Longmeadow, MA, United States of America

Ravindra Mylvaganam