The technique of scanning flow cytometery (SFC) was adopted for measurement of erythrocyte indices: volume, hemoglobin concentration and surface area. The method has been verified on two types of mammalian red cells: human and murine. In order to access distribution of values, precision and stability of inverse algorithm of reconstitution of size and refractive index of microsphere from light scattering angular dependency (indicatrix) have been increased performing analysis of Fourier spectrum of indicatrix. Volume - hemoglobin concentration (V-HbC) map was obtained following well-known procedure of isovolumetric sphering, presented by Technicon Instruments. Additionally new method for measurement of paired distribution of erythrocyte surface area (S) and hemoglobin content (Hb) was implemented via registration of spherical stage in the course of colloid-osmotic hemolysis. Approach to characterization with SFC of native red cells in nonspherical stage is demonstrated.
The kinetics of aluminum (III)-sulfophthalocyanine uptake by human leukocytes was measured with a scanning flow cytometer (SFC) during the initial period of accumulation, 40 min. The individual cells were distinguished by SFC from their light scattering traces. The dye fluorescence in the cells was excited by N2 pulse laser, and the kinetics of the cell distribution on the amount of the accumulated dye was obtained. A mathematical model of endocytosis was applied in order to describe the dynamics of cell distribution in the system during the cellular uptake. The main kinetic parameters of the dye accumulation were evaluated.
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