For the first time, a scanning flow cytometer (SFC) was applied for the analysis of individual particles using angle-resolved light scattering at two wavelengths simultaneously. The SFC was equipped with three lasers with wavelengths of 405, 444 and 660 nm. The performance of dual-wavelength angle-resolved light-scattering (DWARLS) flow cytometry was demonstrated by the analysis of polymer beads, spherized red blood cells (RBCs) and blood platelets. The main advantage of dual-wavelength flow cytometry relates to an increment in the precision of the inverse light-scattering problem solution. The solution allows one to retrieve the characteristics of individual particles by measuring the light-scattering profiles of these particles. DWARLS has allowed us to measure diameters and refractive indices (RIs) of polystyrene beads with record precision: a median error of 12 nm and a few thousandths for diameter and RI, respectively. Analysis of spherized RBCs with DWARLS flow cytometry has provided record precision in the sizing of spherized RBCs, with a median error of 16 nm for diameter, and an adequate value of the specific refraction increment for hemoglobin. By means of DWARLS, we have formed a numerical criterion for shape separation of blood platelets, which can be described by an oblate spheroid model to increase the precision of the distribution parameters of the platelet fractions.
Предметные области OECD FOS+WOS
- 1.03 ФИЗИЧЕСКИЕ НАУКИ И АСТРОНОМИЯ
- 1.03.SY ОПТИКА