V.D. Ustinov

Characterization at the individual cell level and in whole blood samples of shear stress preventing red blood cells aggregation

The aggregation of red blood cells (RBC) is an intrinsic feature of blood that has a strong impact on its microcirculation. For a number of years it has been attracting a great attention in basic research and clinical studies. Here, we study a relationship between the RBC aggregation parameters measured at the individual cell level and in a whole blood sample. The home made optical tweezers were used to measure the aggregating and disaggregating forces for a pair of interacting RBCs, at the individual cell level, in order to evaluate the corresponding shear stresses. The RheoScan aggregometer was used for the measurements of critical shear stress (CSS) in whole blood samples. The correlation between CSS and the shear stress required to stop an RBC pair from aggregating was found. The shear stress required to disaggregate a pair of RBCs using the double channel optical tweezers appeared to be about 10 times higher than CSS. The correlation between shear stresses required to prevent RBCs from aggregation at the individual cell level and in whole blood samples was estimated and assessed quantitatively. The experimental approach developed has a high potential for advancing hemorheological studies.

Study of laser beam scattering by inhomogeneous ensemble of red blood cells in a shear flow

Laser ektacytometry is a technique widely used for measuring the deformability of red blood cells (erythrocytes) in blood samples in vitro. In ektacytometer, a flow of highly diluted suspension of erythrocytes in variable shear stress conditions is illuminated with a laser beam to obtain a diffraction pattern. The diffraction pattern provides information about the shapes (shear-induced elongations) of the cells under investigation. This paper is dedicated to developing the technique of laser ektacytometry so that it would enable one to measure the distribution of the erythrocytes in deformability. We discuss the problem of calibration of laser ektacytometer and test a novel data processing algorithm allowing to determine the parameters of the distribution of erythrocytes deformability. Experimentally, we examined 12 specimens of blood of rats under the action of 4 shear stresses. Analysis of the data shows that in conditions of a limited range of digitizing the diffraction patterns, the measurement errors for the mean deformability, deformability scatter and the skewness of erythrocytes distribution in deformability by our method are respectively 15%, 20% and 20%.

Scattering of a laser beam on a wet blood smear and measurement of red cell size distribution

We report an automated laser system that allows the red cell size distribution to be measured. Experiments are performed on laser light scattering by a suspension of oriented red blood cells (a wet blood smear). Based on an analysis of the angular distribution of light intensity in the diffraction pattern, we have restored the red cell size distribution. The average diameter of a red blood cell is determined with an error of less than 1%, and the spread of red blood cells in size – with an error of about 20%. We discuss the problems of photometry and processing of diffraction patterns, preparing blood samples and data processing algorithms, including methods for solving the inverse scattering problem.