A Novel Methodology to Study Erythrocyte Vesiculation via Oscillatory Shear Flows

We introduce a novel approach to subject erythrocytes to oscillatory shear flows that create tailored shear deformations mimicking the types of deformations that promote vesiculation in splenic flows. This methodology provides an ideal environment to explore a wide variety of metabolic and biochemical effects associated with vesiculation. Deformation details, typical of splenic flow, such as in-folding and implications for membrane/skeleton interaction are demonstrated and quantitatively analyzed through a multiscale numerical model. Moreover, we introduce a theoretical vesiculation model that clearly delineates various fundamental elements that are involved and mediate the vesiculation process and highlights particularly important vesiculation precursors such as areas of membrane/skeleton disruptions that trigger the process. Experiments are carried out in a shear rheometer and we demonstrate via flow cytometry that the deformations we induce on cells do not induce lethal forms of cell damage but do induce vesiculation. This technique is particularly useful in bunch processing of cells necessary for statistical analysis.