Light scatter and total protein signal distribution of platelets by flow cytometry as parameters of size

The use of flow cytometry for determination of platelet size was investigated by measurement of forward-angle light-scatter (FALS) signals and the fluorescent right-angle signals from platelets incubated with the fluorescent protein dye fluorescein isothiocyanate (FITC). Both FALS and fluorescence signals displayed a unimodal log normal pattern of distributions that were almost identical to distributions obtained by resistive particle sizing. As measured on 12 platelet samples from normal individuals, FALS, FITC, and resistive particle size data exhibited a high degree of fit to log normal distributions as shown by the coefficients of determination (R2), which were 0.9962 +/- 0.0035, 0.9966 +/- 0.0056, and 0.9987 +/- 0.0012, respectively. The geometric standard deviation (GSD), reflecting the heterogeneity of the FALS, FITC, and resistive particle size signals, was almost identical: 1.69 +/- 0.03, 1.67 +/- 0.04, and 1.69 +/- 0.02, respectively, for spherical platelets from normal individuals. Platelet FALS signal distributions were compared with Coulter resistive particle size distributions by using platelet samples from 12 normal patients and 27 patients with thrombocytopenia. Significant correlation was found between mean FALS and mean resistive particle size (r = 0.83) and between GSD of FALS and GSD of resistive particle size (r = 0.74) with the platelet samples from the 39 subjects studied. These studies, which document the high degree of correspondence among these three independent measurements of platelet size, based on three entirely different principles, strongly suggest that platelet size is log normal distributed and that the GSD value shown above reflects actual heterogeneity in size. The FALS signal distribution was, however, found to be markedly influenced by changes in platelet shape and internal structure.(ABSTRACT TRUNCATED AT 250 WORDS)