Influence of storage and buffer composition on the mechanical behavior of flowing red blood cells.

On-chip study of blood flow has emerged as a powerful tool to assess the contribution of each component of blood to its overall function. Blood has indeed many functions, from gas and nutrient transport to immune response and thermal regulation. Red blood cells play a central role therein, in particular through their specific mechanical properties, which directly influence pressure regulation, oxygen perfusion, or platelet and white cell segregation toward endothelial walls. As the bloom of in-vitro studies has led to the apparition of various storage and sample preparation protocols, we address the question of the robustness of the results involving cell mechanical behavior against this diversity. The effects of three conservation media (EDTA, citrate, and glucose-albumin-sodium-phosphate) and storage time on the red blood cell mechanical behavior are assessed under different flow conditions: cell deformability by ektacytometry, shape recovery of cells flowing out of a microfluidic constriction, and cell-flipping dynamics under shear flow. The impact of buffer solutions (phosphate-buffered saline and density-matched suspension using iodixanol/Optiprep) are also studied by investigating individual cell-flipping dynamics, relative viscosity of cell suspensions, and cell structuration under Poiseuille flow. Our results reveal that storing blood samples up to 7 days after withdrawal and suspending them in adequate density-matched buffer solutions has, in most experiments, a moderate effect on the overall mechanical response, with a possible rapid evolution in the first 3 days after sample collection.

Migration velocity of red blood cells in microchannels.

The lateral migration of red blood cells (RBCs) in confined channel flows is an important ingredient of microcirculatory hydrodynamics and is involved in the development of a cell free layer near vessel walls and influences the distribution of RBCs in networks. It is also relevant to a number of lab-on-chip applications. This migration is a consequence of their deformability and is due to the combined effects of hydrodynamic wall repulsion and the curvature of the fluid velocity profile. We performed microfluidic experiments with dilute suspensions of RBCs in which the trajectories and migration away from the channel wall are analyzed to extract the mean behavior, from which we propose a generic scaling law for the transverse migration velocity valid in a whole range of parameters relevant to microcirculatory and practical situations. Experiments with RBCs of different mechanical properties (separated by density gradient sedimentation or fixed with glutaraldehyde) show the influence of this parameter which can induce significant dispersion of the trajectories.