Red blood cells (RBCs) are the most abundant cells in human blood. Remarkably RBCs deform and bridge together to form aggregates under very low shear rates. The theory and mechanics behind aggregation are, however, not yet completely understood. The main objective of this work is to quantify and characterize RBC aggregates in order to enhance the current understanding of the non-Newtonian behaviour of blood in microcirculation. Suspensions of human blood were flowed and observed in vitro in poly-di-methyl-siloxane (PDMS) microchannels to characterize RBC aggregates. These microchannels were fabricated using standard photolithography methods. Experiments were performed using a micro particle image velocimetry (μPIV) system for shear rate measurements, coupled with a high-speed camera for flow visualization. RBC aggregate sizes were quantified in controlled and measurable shear rate environments for 5, 10 and 15% hematocrit. Aggregate sizes were determined using image processing techniques, while apparent viscosity was measured using optical viscometry. For the samples suspended at 5% H, aggregate size was not strongly correlated with shear rate. For the 10% H suspensions, in contrast, lowering the shear rate below 10 s-1 resulted in a significant increase of RBC aggregate sizes. The viscosity was found to increase with decreasing shear rate and increasing hematocrit, exemplifying the established non-Newtonian shear-thinning behaviour of blood. Increase in aggregation size did not translate into a linear increase of the blood viscosity. Temperature was shown to affect blood viscosity as expected, however, no correlation for aggregate size with temperature was observed. Non-Newtonian parameters associated with power law and Carreau models were determined by fitting the experimental data and can be used towards the simple modeling of blood’s non-Newtonian behaviour in microcirculation. This work establishes a relationship between RBC aggregate sizes and corresponding shear rates and one between RBC aggregate sizes and apparent blood viscosity at body and room temperatures, in a microfluidic environment for low hematocrit. Effects of hematocrit, shear rate, viscosity and temperature on RBC aggregate sizes have been quantified.

The intent of this paper is to investigate the application of a pre-processing method previously validated on glycerol to blood flows in microchannels and to compare the accuracy of results obtained when applied to a non-homogeneous fluid such as blood with results from previously applied processing methods for blood data. Comparisons of common processing methods are desired for a clear measure of accuracy in order to make recommendations for various flows. It is hypothesized that increasing the correlation window overlap improves the profile prediction. The amount of correlation window overlap and window shape in the processing of data have a significant effect on the results. Image pre-processing is explored to improve the correlation using the ‘image overlapping’ which is extended to the case of blood and the blood-specific pre-processing ‘base-clipping’ or ‘thresholding’ technique currently applied to blood. Both pre-processing methods are tested with multiple processing methods for two channel geometries: a straight rectangular channel and a Y-channel resulting in a controlled shear flow. The resulting profiles and calculations demonstrate that ‘image-overlapping’ is found to achieve a profile closer to the predicted theoretical profile than current blood pre-processing methods when both are applied to the same set of data and both are superior to conventional cross-correlation on its own. In all cases, pre-processing decreases the smoothness of the predicted profile. The use of ‘image-overlapping’ is shown to have greater accuracy when calculating the shear rate at the wall of the channel as well.

The purpose of this paper is to design a microfluidic apparatus capable of providing controlled flow conditions suitable for red blood cell (RBC) aggregation analysis. The linear velocity engendered from the controlled flow provides constant shear rates used to qualitatively analyze RBC aggregates. The design of the apparatus is based on numerical and experimental work. The numerical work consists of 3D numerical simulations performed using a research computational fluid dynamics (CFD) solver, Nek5000, while the experiments are conducted using a microparticle image velocimetry system. A Newtonian model is tested numerically and experimentally, then blood is tested experimentally under several conditions (hematocrit, shear rate, and fluid suspension) to be compared to the simulation results. We find that using a velocity ratio of 4 between the two Newtonian fluids, the layer corresponding to blood expands to fill 35% of the channel thickness where the constant shear rate is achieved. For blood experiments, the velocity profile in the blood layer is approximately linear, resulting in the desired controlled conditions for the study of RBC aggregation under several flow scenarios.

Blood, as a non-Newtonian biofluid, represents the focus of numerous studies in the hemorheology field. Blood constituents include red blood cells, white blood cells and platelets that are suspended in blood plasma. Due to the abundance of the RBCs (40% to 45% of the blood volume), their behavior dictates the rheological behavior of blood especially in the microcirculation. At very low shear rates, RBCs are seen to assemble and form entities called aggregates, which causes the non-Newtonian behavior of blood. It is important to understand the conditions of the aggregates formation to comprehend the blood rheology in microcirculation. The protocol described here details the experimental procedure to determine quantitatively the RBC aggregates in microcirculation under constant shear rate, based on image processing. For this purpose, RBCsuspensions are tested and analyzed in 120 x 60 µm poly-dimethyl-siloxane (PDMS) microchannels. The RBC-suspensions are entrained using a second fluid in order to obtain a linear velocity profile within the blood layer and thus achieve a wide range of constant shear rates. The shear rate is determined using a micro Particle Image Velocimetry (µPIV) system, while RBC aggregates are visualized using a high speed camera. The videos captured of the RBC aggregates are analyzed using image processing techniques in order to determine the aggregate sizes based on the images intensities. Video LinkThe video component of this article can be found at

This work aims to develop a method for quantitatively analyzing red blood cell (RBC) aggregates under controlled flow conditions. Images of experiments are captured and processed in order to quantify aggregation. The experimental setup consists of RBC suspensions in blood plasma entrained by a phosphate-buffered saline solution in a 110 × 60 µm polydimethylsiloxane microchannel. The experiments are performed by varying the hematocrit (5, 10, and 15%) and the flow rate (Q = 5 and 10 µl/hr) in order to observe the effect of shear rate on RBC aggregation. Microchannel dimensions as well as fluid flow rates are determined using numerical simulations. The flow is visualized using a highspeed camera coupled to a micro particle image velocimetry system. Videos obtained with the high-speed camera are processed using a MATLAB program, with each frame analyzed separately. RBC aggregates are detected based on the image intensities and the connectivity between RBCs, using image processing techniques. The average aggregate size and distribution of RBCs for various aggregate sizes are determined for each of the shear rates and hematocrits. These aggregates are shown to be larger at low flow rates where the shear rate is small. Results from tests performed at high hematocrits also show larger RBC aggregates.