Characterization of the Normoblast Population in β‐Thalassaemic Blood by Rapid‐Flow Cytofluorometry

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Abstract
Samples of peripheral blood from patients with .beta.-thalassemia major which contained significant numbers of nucleated normoblasts were stained with acridine orange and analyzed with rapid-flow cytofluorometry. The pyknotic normoblast-nuclei gave less green DNA fluorescence than the (diploid) leukocytes and constituted a separate, distinct subpopulation. Mean values of the fluorescence intensities and standard deviations as displayed by multichannel analyses gave a numerical value for normoblasts with regard to their maturation stages. These mean values correlated with the differential counts of early and late normoblasts in the light microscope under rigidly standardized conditions. Rapid-flow cytofluorometry provides an objective and quantitative way to monitor and define peripheral blood normoblast populations as a measure of the severity of erythropoietic stress.