Annie Viallat

Summary: There is a number of publications on red blood cell deformability, that is, on the remarkable cell ability to change its shape in response to an external force and to pass through the narrowest blood capillaries and splenic sinuses. Cell deformability is postulated to be a major determinant of impaired perfusion, increase of blood viscosity, and occlusion in microvessels. Current deformability tests like ektacytometry measure global parameters, related to shape changes at the whole cell scale. Despite strong advances in our understanding of the molecular organization of red blood cells, the relationships between the rheology of each element of the cell composite structure, the global deformability tests, and the cell behavior in microflows are still not elucidated. This review describes recent advances in the description of the dynamics of red blood cells in shear flow and in the mechanistic understanding of this dynamics at the scale of the constitutive rheological and structural elements of the cell. These developments could open up new horizons for the determination of red blood cell mechanical parameters by analyzing their motion under low shear flows.

At the cellular scale, blood fluidity and mass transport depend on the dynamics of red blood cells in blood flow, specifically on their deformation and orientation. These dynamics are governed by cellular rheological properties, such as internal viscosity and cytoskeleton elasticity. In diseases in which cell rheology is altered genetically or by parasitic invasion or by changes in the microenvironment, blood flow may be severely impaired. The nonlinear interplay between cell rheology and flow may generate complex dynamics, which remain largely unexplored experimentally. Under simple shear flow, only two motions, “tumbling” and “tank-treading,” have been described experimentally and relate to cell mechanics. Here, we elucidate the full dynamics of red blood cells in shear flow by coupling two videomicroscopy approaches providing multidirectional pictures of cells, and we analyze the mechanical origin of the observed dynamics. We show that contrary to common belief, when red blood cells flip into the flow, their orientation is determined by the shear rate. We discuss the “rolling” motion, similar to a rolling wheel. This motion, which permits the cells to avoid energetically costly deformations, is a true signature of the cytoskeleton elasticity. We highlight a hysteresis cycle and two transient dynamics driven by the shear rate: an intermittent regime during the “tank-treading-to-flipping” transition and a Frisbee-like “spinning” regime during the “rolling-to-tank-treading” transition. Finally, we reveal that the biconcave red cell shape is highly stable under moderate shear stresses, and we interpret this result in terms of stress-free shape and elastic buckling.

We have used an original analysis of reflection interference contrast microscopy (RICM) to detect an ultra-weak specific interaction between a glycolipid vesicle and a lectin-coated substrate. The membrane height fluctuations in the contact zone are observed with a high illumination aperture; the membrane profile and the membrane-substrate distance are quantitatively determined using the new analysis, which accounts for multiple interfaces and multiple incidence rays. We showed that this refined version of RICM theory is necessary, specifically in the case of intermediate membranesubstrate distance (30 nm) and helped to discriminate between the ultra-weak interaction and pure gravitational sedimentation

We show that the motion of individual red blood cells in an oscillating moderate shear flow is described by a nonlinear system of three coupled oscillators. Our experiments reveal that the cell tank~treads and tumbles either in a stable way with synchronized cell inclination, membrane rotation and hydrodynamic oscillations, or in an irregular way, very sensitively to initial conditions. By adapting our model described previously, we determine the theoretical diagram for the red cell motion in a sinusoidal flow close to physiological shear stresses and flow variation frequencies and reveal large domains of chaotic motions. Finally, fitting our observations allows a characterization of cell viscosity and membrane elasticity.

We have used an original analysis of reflection interference contrast microscopy (RICM) to detect an ultra-weak specific interaction between a glycolipid vesicle and a lectin-coated substrate. The membrane height fluctuations in the contact zone are observed with a high illumination aperture; the membrane profile and the membrane-substrate distance are quantitatively determined using the new analysis, which accounts for multiple interfaces and multiple incidence rays. We showed that this refined version of RICM theory is necessary, specifically in the case of intermediate membrane-substrate distance (∼30 nm) and helped to discriminate between the ultra-weak interaction and pure gravitational sedimentation