Method for hematology analysis

摘要

A method whereby one or more fluorescent dyes are used to bind and stain nucleic acids in certain blood cells, such as, for example, white blood cells, nucleated red blood cells, and reticulocytes, and to induce fluorescent emissions upon excitation of photons from a given source of light, such as, for example, a laser, at an appropriate wavelength. More particularly, this invention provides a method whereby a fluorescent trigger is used in a data collection step for collecting events that emit strong fluorescence, in order to separate white blood cells and nucleated red blood cells from red blood cells and platelets without the need for using a lysing agent.

主权项

1. A method for counting cells in a sample of whole blood by means of an automated hematology analyzer, said method comprising the steps of:
(a) diluting a sample of whole blood with a diluent, wherein the sample comprises a plurality of white blood cells, nucleated red blood cells, red blood cells, platelets, and reticulocytes; (b) contacting the sample with at least one fluorescent dye that specifically binds to and stains one or more nucleic acids in the white blood cells, the nucleated red blood cells, and/or the reticulocytes in the sample; (c) passing the sample through the hematology analyzer to generate a plurality of events, wherein each event comprises a plurality of light scattering signals and a fluorescence emission signal generated by a cell in the sample; (d) separating the events into two data blocks, prior to counting the events, using a fluorescent trigger configured in the hematology analyzer, wherein the first data block includes events having a fluorescence emission signal that is below the fluorescent trigger, and the second data block includes events having a fluorescence emission signal that is above the fluorescent trigger; (e) analyzing the events in the first data block to count the number of red blood cells, platelets, and reticulocytes in the sample; (f) analyzing the events in the second data block to count the number of white blood cells and nucleated red blood cells in the sample.