Open AccessOptimization of flow‐cytometric discrimination between reticulocytes and erythrocytes
Author(s) -
Schmitz Franz J.,
Werner Eckhard
Publication year - 1986
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990070508
Subject(s) - acridine orange , reticulocyte , fixative , flow cytometry , fluorescence , microbiology and biotechnology , cytometry , hemolysis , chromatography , biology , staining , chemistry , biochemistry , immunology , genetics , physics , quantum mechanics , messenger rna , gene
An automatized technique to count reticulocytes by means of flow cytometry is described. Blood samples were stained by the fluorescent dye acridine orange without the use of fixative. Scatter and red fluorescence of the blood cells were measured in a flow cytometer. A discrimination between reticulocytes and erythrocytes was only achieved by using logarithmic amplification. The discrimination was better in peak mode than in area mode. The optimum dye concentration was 0.5 mg/liter acridine orange. At lower dye concentrations, not all reticulocytes were measured, whereas at higher dye concentrations the degree of discrimination between reticulocytes and erythrocytes decreased. There was a suitable discrimination between reticulocytes and erythrocytes. The reticulocyte numbers were scored by flow cytometry as well as by microscope for blood samples with 0.1–14% reticulocytes. The correlation between both methods was close.