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. . Its fluidity strongly depends on its behavior in flow, which is a key factor of proper tissue perfusion. At the cellular scale, blood flow behavior is affected primarily by the RBC response to the hydrodynamic stress in terms of cell orientation relative to the flow direction and of cell deformation. For example, on one hand, at low shear rates, similar cell orientations may favor the formation of stacks (rouleaux) (1) of RBCs, like rolls of coins, which increases blood viscosity. On the other hand, at high shear rates, the individualization of RBCs, their alignment, and their stretching in the flow (2) decrease blood viscosity (3). The orientation and the deformation in flow of RBCs are governed by their rheological properties. They result from the viscoelastic contributions of all components of the cell composite structure. Moreover, RBC rheological properties also depend on the microenvironment and on metabolic functionality (4). Both local and systemic disturbances of homeostasis (in diabetes mellitus, hypertension) have the potential to induce RBC rheological alterations and consequently to impair blood circulation. It therefore is crucial to understand the relationships between the rheological properties of RBCs and their orientation and deformation in flow. This question is far from trivial because even in a simple shear flow, RBCs present a v...

The concept of using stimuli-responsive hydrogels to actuate fluids in microfluidic devices is particularly attractive, but limitations, in terms of spatial resolution, speed, reliability and integration, have hindered its development during the past two decades. By patterning and grafting poly(N-isopropylacrylamide) PNIPAM hydrogel films on plane substrates with a 2 μm horizontal resolution and closing the system afterward, we have succeeded in unblocking bottlenecks that thermo-sensitive hydrogel technology has been challenged with until now. In this paper, we demonstrate, for the first time with this technology, devices with up to 7800 actuated micro-cages that sequester and release solutes, along with valves actuated individually with closing and opening switching times of 0.6 ± 0.1 and 0.25 ± 0.15 s, respectively. Two applications of this technology are illustrated in the domain of single cell handling and the nuclear acid amplification test (NAAT) for the Human Synaptojanin 1 gene, which is suspected to be involved in several neurodegenerative diseases such as Parkinson's disease. The performance of the temperature-responsive hydrogels we demonstrate here suggests that in association with their moderate costs, hydrogels may represent an alternative to the actuation or handling techniques currently used in microfluidics, that are, pressure actuated polydimethylsiloxane (PDMS) valves and droplets.

An analytical model was proposed by Keller and Skalak in 1982 to understand the motion of red blood cells in shear flow. The cell was described as a fluid ellipsoid of fixed shape. This model was extended in 2007 to introduce shear elasticity of the red blood cell membrane. Here, this model is further extended to take into account that the cell discoid shape physiologically observed is not a stress-free shape. The model shows that spheroid stress-free shapes allow us to fit the experimental data with the values of shear elasticity typical to that found with micropipette and optical tweezer experiments. In the range of moderate shear rates (for which RBCs keep their discoid shape) this model enables us to quantitatively determine (i) an effective cell viscosity, which combines membrane and hemoglobin viscosities and (ii) an effective shear modulus of the membrane that combines the shear modulus and the stress-free shape. This model can also be used to determine RBC mechanical parameters not only in the tanktreading regime when cells are suspended in medium of high viscosity but also in the tumbling regime characteristic of cells suspended in media of low viscosity. In this regime, a transition is predicted between a rigid-like tumbling motion and a fluid-like tumbling motion above a critical shear rate, which is directly related to the mechanical parameters of the cell.