Determination of white blood cell differential and reticulocyte counts

摘要

A method of identifying target subpopulations of nucleated cells (34) contained in a quiescent sample of anticoagulated whole blood which is mixed with a fluorescent dye, wherein the blood sample is diluted by no more than about 1: 1; wherein said whole blood sample anticoagulated dispersed is dispersed in a viewing chamber (14) including a plurality of regions of different thickness, comprising: a first region (A) having a first thickness across the plane between two seven microns which allows the inclusion of individual red blood cells (32) and / or monolayers (31) of red blood cells; and a second region (B) having a second thickness through the plane, between seven and Forty-micron; said method comprising: a) the exploration phase of selected fields of view (1, 3, 5) in those regions with an optical scanning instrument (54); b) the step of lighting of said selected fields of view (1, 3, 5) with a light source of one or more preselected wavelengths that are functional to cause said fluorescent dye produces fluorescent emissions in one or more lengths known wave, the said emission characteristics of said target subpopulations of nucleated cells; c) because the characteristic fluorescence of cells to said known wavelengths including 530 nm, 585 nm and 660 nm when excited at the preselected length in the range of 485 nm, and its fluorescence at said another wavelength known 610 nm when the preselected wavelength in the range of 560 nm is excited, the step of analyzing said emissions to said known wavelengths to identify and distinguish said target nucleated cell subpopulations; wherein the chamber (14) having opposed containment walls of the sample (4, 8), and the term "through plane" refers to a line of sight corresponding to the shortest distance between the walls (4 , 8) in a given chamber (14) region.