Measurement of Mammalian Erythrocyte Indices from Light Scattering with Scaning Flow Cytometer

摘要

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.