Migration Of Red Blood Cells Research Articles

Advanced design of indented-standing surface acoustic wave (i-SSAW) device for platelet-derived microparticle separation from whole blood

Isolating submicron microvesicles (MVs) from blood has become important for clinical diagnosis and researches. Advanced design of indented-standing surface acoustic wave (i-SSAW)-based device with a U-shaped microchannel is proposed for extracting platelet-derived microparticles (PDMPs) from whole blood. Unlike the previous acoustic devices, the cross-sectional shape of the microchannel was manipulated to enhance the device performance, rather than the patterned interdigitated transducers. Cross-sectional shape-dependent device performance was evaluated using rectangular channels with different heights and U-shaped channel with whole blood sample with 40 % hematocrit. Effect of the flow rates on acoustic migration of red blood cells (RBCs) was examined at 20 V fixed voltage. In addition, RBC and platelet removal was examined using different applied voltages for i-SSAW operation. Finally, high-purity (approximately 100 %) and high-recovery (approximately 55.6 %) platelet-poor plasma was separated from whole blood at 30 μL/min without platelet activation.

Electromagnetism - Properties of Erythrocytes

At the onset of blood flow, red blood cells (RBCs) align along the central plane of the vessels. In capillaries, RBCs transform from a biconcave disk into a parachute shape. As blood transitions from the arterial to the venous end, the hemoglobin in erythrocytes alters its magnetic susceptibility. Within approximately 0.6-0.8 seconds, oxygen is displaced from RBCs through diffusion. This study explores the fundamentals and interconnections of these processes.
 
 Blood samples from 35 different healthy individuals were analyzed. The research examined magnetic field induction in ferromagnetic toroids, formed by the alternative electric field with a square wave signal, and studied the AC in the secondary coil - a tube filled with blood. This study discusses the impact of RBC geometry and hemoglobin allosteric transitions on electric signal generation and its relevance to cellular metabolic activity in the body.
 
 The findings suggest that the AC field, originating from the heart's rotational dipole, can generate a magnetic field in RBCs, facilitating the allosteric transformations of hemoglobin. Hemoglobin's thermoelastic expansion and magnetostriction cause biconcave membrane oscillations at ultrasound frequencies. The resulting electroacoustic wave rotates charges at the cell's z-potential area, aids RBC migration into the flow plane, and enhances trans capillary diffusion of substances.
 
 An electroacoustic standing wave emerges between the oscillating RBCs, coinciding with the wavenumber of externally penetrating infrared light. The synergic influence on hemoglobin in capillaries causes the RBC membrane to create a temporally frequency-modulated wave, carrying resonance molecular frequencies. This wave regulates biochemical processes within and outside body cells.

Modeling of three-dimensional blood flow in microchannels using a two-fluid method

This work presents a novel two-fluid method based on our recently proposed viscosity model for red blood cells (RBCs)—for simulating three-dimensional (3D) blood flow in a microchannel of dimension comparable to the diameter of red blood cells and larger. Toward this, whole blood is assumed as a suspension of red blood cells in blood plasma, with each phase considered as interpenetrating continua having its separate mass and momentum conservation equations. The proposed approach-based performance study is presented after comprehensively validating it with experimental data for blood flow in a uniform, sudden expansion-constriction, and Y-shaped bifurcated rectangular microchannels over—an extensive range of size (25–330 μm), flow rates (11.8 μl/h–30 ml/h), and inlet hematocrit (0%–45%). The proposed approach effectively captures significant biophysical and biomechanical insights into blood flow. It highlights a migration of red blood cells toward the center of the microchannel and the formation of a cell-free layer near the wall. Notably, with the introduction of constriction and expansion in the microchannel, it predicts a fivefold enhancement of the cell-free layer. The Fahraeus and Fahraeus–Lindquist effects are also demonstrated in microchannels, with less than 300 μm characteristic dimensions. These findings are consistent with experimental evidence. In addition to experimentally evident phenomena, our simulations unveil several additional flow phenomena and features of blood flow in the microchannel. It is observed that the presence of confluence (merging flow) is more disturbing to the blood flow than the presence of diverging bifurcations (splitting flow). Furthermore, after the confluence, velocity profiles exhibit a local peak that persists up to the microchannel outlet. Primary contribution of this work lies in the proposal of a two-fluid method for simulating 3D blood flow in complex geometries. This approach provides a comprehensive understanding of blood flow dynamics in microchannels and can be applied to optimize dimensions and geometries during the initial phases of plasma separation microdevices development.

Migration Study of Dielectrophoretically Manipulated Red Blood Cells in Tapered Aluminium Microelectrode Array: A Pilot Study.

Dielectrophoresis (DEP) is one of the microfluid-based techniques that can manipulate the red blood cells (RBC) for blood plasma separation, which is used in many medical screening/diagnosis applications. The tapered aluminium microelectrode array (TAMA) is fabricated for potential sensitivity enhancement of RBC manipulation in lateral and vertical directions. In this paper, the migration properties of dielectrophoretically manipulated RBC in TAMA platform are studied at different peak-to-peak voltage (Vpp) and duration supplied onto the microelectrodes. Positive DEP manipulation is conducted at 440 kHz with the RBC of 4.00 ± 0.2 µm average radius attracted to the higher electric field intensity regions, which are the microelectrodes. High percentage of RBC migration occurred at longer manipulation time and high electrode voltage. During DEP manipulation, the RBC are postulated to levitate upwards, experience the electro-orientation mechanism and form the pearl chains before migrating to the electrodes. The presence of external forces other than the dielectrophoretic force may also affect the migration response of RBC. The safe operating limit of 10 Vpp and manipulation duration of ≤50 s prevent RBC rupture while providing high migration percentage. It is crucial to define the safe working region for TAMA devices that manipulate small RBC volume (~10 µL).

Influence of the vessel wall geometry on the wall-induced migration of red blood cells.

The geometry of the blood vessel wall plays a regulatory role on the motion of red blood cells (RBCs). The overall topography of the vessel wall depends on many features, among which the endothelial lining of the endothelial surface layer (ESL) is an important one. The endothelial lining of vessel walls presents a large surface area for exchanging materials between blood and tissues. The ESL plays a critical role in regulating vascular permeability, hindering leukocyte adhesion as well as inhibiting coagulation during inflammation. Changes in the ESL structure are believed to cause vascular hyperpermeability and entrap immune cells during sepsis, which could significantly alter the vessel wall geometry and disturb interactions between RBCs and the vessel wall, including the wall-induced migration of RBCs and the thickening of a cell-free layer. To investigate the influence of the vessel wall geometry particularly changed by the ESL under various pathological conditions, such as sepsis, on the motion of RBCs, we developed two models to represent the ESL using the immersed boundary method in two dimensions. In particular, we used simulations to study how the lift force and drag force on a RBC near the vessel wall vary with different wall thickness, spatial variation, and permeability associated with changes in the vessel wall geometry. We find that the spatial variation of the wall has a significant effect on the wall-induced migration of the RBC for a high permeability, and that the wall-induced migration is significantly inhibited as the vessel diameter is increased.

Inertial migration of red blood cells under a Newtonian fluid in a circular channel

We present a numerical analysis of the lateral movement and equilibrium radial positions of red blood cells (RBCs) with major diameter 8 $\mathrm {\mu }$ m under a Newtonian fluid in a circular channel with 50 $\mathrm {\mu }$ m diameter. Each RBC, modelled as a biconcave capsule whose membrane satisfies strain-hardening characteristics, is simulated for different Reynolds numbers $Re$ and capillary numbers $Ca$ , the latter of which indicates the ratio of the fluid viscous force to the membrane elastic force. The effects of initial orientation angles and positions on the equilibrium radial position of an RBC centroid are also investigated. The numerical results show that depending on their initial orientations, RBCs have bistable flow modes, so-called rolling and tumbling motions. Most RBCs have a rolling motion. These stable modes are accompanied by different equilibrium radial positions, where tumbling RBCs are further away from the channel axis than rolling ones. The inertial migration of RBCs is achieved by alternating orientation angles, which are affected primarily by the initial orientation angles. Then the RBCs assume the aforementioned bistable modes during the migration, followed by further migration to the equilibrium radial position at much longer time periods. The power (or energy dissipation) associated with membrane deformations is introduced to quantify the state of membrane loads. The energy expenditures rely on stable flow modes, the equilibrium radial positions of RBC centroids, and the viscosity ratio between the internal and external fluids.

In vivo real-time red blood cell migration and microcirculation flow synergy imaging-surveyed thrombolytic therapy with iron-oxide complexes.

Nanotherapeutics as a nascent method has attracted widely interest on the treatment of thrombosis. However, due to the limited temporal and spatial resolution of conventional imaging modalities, the dynamic visualization the thrombogenesis and evaluation of the effect of thrombolytic drugs are facing severely difficulties in vivo. In addition, the development of high targeting, short circulation time, and small size thrombolysis nanotherapeutics agents requires further research. Herein, we report a synergy imaging modality that combining a label-free capillary microscopy and laser speckle microcirculation imaging, which realized dynamic visualization of single red blood cell migration and large-field dynamic blood flow. In this work, we investigated the red blood cells migration and blood flow velocity response before and after treated through introducing a functional nano-thrombolytics, iron-oxide complexes coated urokinase (IPN@UK) on an orthotopic animal model in vivo. The functionalized IPN@UK nanocomposites exhibited outstanding thrombolysis effect. Significantly, whole-course changes, including red blood cell activity, complex thrombolytic therapeutics, were well surveilled and evaluated using dual-modality combining imaging strategy. These results show this synergy imaging strategy not only can achieve multiscale non-invasive visualization of dynamic thrombus events in real-time, but also can quantify hemodynamics information of thrombus. Our study demonstrates the potential of this synergy imaging method, which for early detection of thrombus, evaluation of the effect of drug thrombolysis, developing the thrombolytic drugs, and imaging-guide thrombolytic therapy in living systems.

Resolved CFD-DEM simulation of blood flow with a reduced-order RBC model

Mathematical modeling of the blood flow with a resolved description of biological cells mechanics such as red blood cell (RBC) has been a challenge in the past decades as it involves physical complexities and demands high computational costs. In the present study, we propose an approach for efficient simulation of blood flow with several suspended RBCs. In this approach, we employ our previously proposed reduced-order model for deformable particles (Nair et al. in Comput Part Mech 7:593–601, 2020) to mimic the mechanical behavior of an individual RBC as a cluster of overlapping spheres interconnected by flexible mathematical bonds. This discrete element method-based model is then coupled with a fluid flow solver using the immersed boundary method with continuous forcing in the context of computational fluid dynamics-discrete element method (CFD-DEM) coupling. The present computational method is tested with a couple of validation cases in which the single RBC dynamics, as well as the blood flow with several RBCs, were tested in comparison with existing literature date. First, the RBC deformation index in shear flow at different shear rates is studied with a good accuracy. Then, the blood flow in micro-tubes of different diameters and hematocrits was simulated. The key characteristics of blood flow such as cell-free layer (CFL) thickness, Fahraeus effect and the relative apparent viscosity are used as the validation metrics. The proposed approach can predict the formation of the migration of RBC toward the tube center-line and the CFL thickness in good agreement with previous measurement and simulations. Furthermore, the model is employed to study the CFL enhancement for plasma separation based on channel constriction. The simulation results compute the CFL thickness downstream of the channel constriction in good agreement with the experiments in a wide range of flow rates and constriction lengths. The original contribution of this study lies in proposing an efficient resolved CFD-DEM simulation method for blood flows with many RBCs which can be employed for numerical investigation of bio-microfluidic applications.

Axial and Nonaxial Migration of Red Blood Cells in a Microtube.

Human red blood cells (RBCs) are subjected to high viscous shear stress, especially during microcirculation, resulting in stable deformed shapes such as parachute or slipper shape. Those unique deformed RBC shapes, accompanied with axial or nonaxial migration, cannot be fully described according to traditional knowledge about lateral movement of deformable spherical particles. Although several experimental and numerical studies have investigated RBC behavior in microchannels with similar diameters as RBCs, the detailed mechanical characteristics of RBC lateral movement—in particular, regarding the relationship between stable deformed shapes, equilibrium radial RBC position, and membrane load—has not yet been fully described. Thus, we numerically investigated the behavior of single RBCs with radii of 4 m in a circular microchannel with diameters of 15 m. Flow was assumed to be almost inertialess. The problem was characterized by the capillary number, which is the ratio between fluid viscous force and membrane elastic force. The power (or energy dissipation) associated with membrane deformations was introduced to quantify the state of membrane loads. Simulations were performed with different capillary numbers, viscosity ratios of the internal to external fluids of RBCs, and initial RBC centroid positions. Our numerical results demonstrated that axial or nonaxial migration of RBC depended on the stable deformed RBC shapes, and the equilibrium radial position of the RBC centroid correlated well with energy expenditure associated with membrane deformations.

Continuum Scale Non Newtonian Particle Transport Model for Hæmorheology

We present a continuum scale particle transport model for red blood cells following collision arguments, in a diffusive flux formulation. The model is implemented in FOAM, in a framework suitable for haemodynamics simulations and adapted to multi-scaling. Specifically, the framework we present is able to ingest transport coefficient models to be derived, prospectively, from complimentary but independent meso-scale simulations. For present purposes, we consider modern semi-mechanistic rheology models, which we implement and test as proxies for such data. The model is verified against a known analytical solution and shows excellent agreement for high quality meshes and good agreement for typical meshes as used in vascular flow simulations. Simulation results for different size and time scales show that migration of red blood cells does occur on physiologically relevany timescales on small vessels below 1 mm and that the haematocrit concentration modulates the non-Newtonian viscosity. This model forms part of a multi-scale approach to haemorheology and model parameters will be derived from meso-scale simulations using multi-component Lattice Boltzmann methods. The code, haemoFoam, is made available for interested researchers.

Spatiotemporal Dynamics of Dilute Red Blood Cell Suspensions in Low-Inertia Microchannel Flow

Microfluidic technologies are commonly used for the manipulation of red blood cell (RBC) suspensions and analyses of flow-mediated biomechanics. To enhance the performance of microfluidic devices, understanding the dynamics of the suspensions processed within is crucial. We report novel, to our knowledge, aspects of the spatiotemporal dynamics of RBC suspensions flowing through a typical microchannel at low Reynolds number. Through experiments with dilute RBC suspensions, we find an off-center two-peak (OCTP) profile of cells contrary to the centralized distribution commonly reported for low-inertia flows. This is reminiscent of the well-known “tubular pinch effect,” which arises from inertial effects. However, given the conditions of negligible inertia in our experiments, an alternative explanation is needed for this OCTP profile. Our massively parallel simulations of RBC flow in real-size microfluidic dimensions using the immersed-boundary-lattice-Boltzmann method confirm the experimental findings and elucidate the underlying mechanism for the counterintuitive RBC pattern. By analyzing the RBC migration and cell-free layer development within a high-aspect-ratio channel, we show that such a distribution is co-determined by the spatial decay of hydrodynamic lift and the global deficiency of cell dispersion in dilute suspensions. We find a cell-free layer development length greater than 46 and 28 hydraulic diameters in the experiment and simulation, respectively, exceeding typical lengths of microfluidic designs. Our work highlights the key role of transient cell distribution in dilute suspensions, which may negatively affect the reliability of experimental results if not taken into account.

A Two-Zone Shear-Induced Red Blood Cell Migration Model for Blood Flow in Microvessels

The Fahraeus and Fahraeus-Lindqvist effects are both associated with the concentration of red blood cells (RBCs) in the core region of microvessels. The annular region is a cell-free layer. Blood flow dynamics and both effects are related to the hematocrit level profile. The aim is to propose a model for blood flow in microvessels that is not compute intensive like many other models such as those using finite element methods. Modeling blood flow requires solving for both the hematocrit level and velocity profiles as blood viscosity depends on the hematocrit level. The two-zone shear-induced model for blood flow is adopted while including an annular cell-free layer, as in the marginal zone theory and in consistency with experimental observations. In the core region, the hematocrit level is not considered to be uniform, and the concentration and viscous fluxes are equal in magnitude and opposite in directions in the fully developed velocity and concentration profiles case. The momentum and hematocrit balance equations are solved. Both analytical and numerical solutions for the velocity and hematocrit level profiles are determined. The numerical results are found to exactly match the analytical solutions, and to be in very good agreement with published experimental data for the cell-free layer thickness, the velocity profile, and the hematocrit ratio.

The Fåhræus-Lindqvist effect in small blood vessels: how does it help the heart?

The Fåhræus-Lindqvist effect is usually explained from a physical point of view with the so-called Haynes' marginal zone theory, i.e., migration of red blood cells (RBCs) to a core layer surrounded by an annular RBCs-free plasma layer. In this paper we show that the marginal layer, though causing a substantial reduction in flow resistance and increasing discharge, does not reduce the rate of energy dissipation. This fact is not surprising if one considers the electric analog of the flow in a vessel: a resistance reduction increases both the current intensity (i.e., the discharge) and the energy dissipation. This result is obtained by considering six rheological models that relate the blood viscosity to hematocrit (volume fraction occupied by erythrocytes). Some physiological implications are discussed.

EFFECT OF STENOSIS SEVERITY ON SHEAR-INDUCED DIFFUSION OF RED BLOOD CELLS IN CORONARY ARTERIES

In large blood vessels, migration of red blood cells (RBCs) affects the concentration of platelets and the transport of oxygen to the arterial endothelial cells. In this work, we investigate the locations where hydrodynamic diffusion of RBCs occurs and the effects of stenosis severity on shear-induced diffusion (SID) of RBCs, concentration distribution and wall shear stress (WSS). For the first time, multiphase mixture theory approach with Phillips shear-induced diffusive flux model coupled with Quemada non-Newtonian viscosity model has been applied to numerically simulate the RBCs macroscopic behavior in four different degrees of stenosis (DOS) geometries, viz., 30%, 50%, 70% and 85%. Considering SID of RBCs, the calculated average WSS increased by 77.70% which emphasises the importance of SID in predicting hemodynamic parameters. At the stenosis throat, it was observed that 85% DOS model had the lowest concentration of RBCs near the wall and highest concentration at the center. For the stenosis models with 70% and 85% DOS, the RBC lumen wall concentration at the distal section of stenosis becomes inhomogeneous with the maximum fluctuation of 1.568%. Finally, the wall regions with low WSS and low RBC concentrations correlate well with the atherosclerosis sites observed clinically.

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.

Dynamics of blood flow: modeling of Fåhraeus and Fåhraeus-Lindqvist effects using a shear-induced red blood cell migration model.

Blood flow in micro capillaries of diameter approximately 15-500μm is accompanied with a lower tube hematocrit level and lower apparent viscosity as the diameter decreases. These effects are termed the Fåhraeus and Fåhraeus-Lindqvist effects, respectively. Both effects are linked to axial accumulation of red blood cells. In the present investigation, we extend previous works using a shear-induced model for the migration of red blood cells and adopt a model for blood viscosity that accounts for the suspending medium viscosity and local hematocrit level. For fully developed hematocrit profiles (i.e., independent of axial location), the diffusion fluxes due to particle collision frequency and viscosity gradients are of equal magnitude and opposite directions. The ratio of the diffusion coefficients for the two fluxes affects both the Fåhraeus and Fåhraeus-Lindqvist effects and is found related to the capillary diameter and discharge hematocrit using a well-known data-fit correlation for apparent blood viscosity. The velocity and hematocrit profiles were determined numerically as functions of radial coordinate, tube diameter, and discharge hematocrit. The velocity profile determined numerically is consistent with the derived analytical expression and the results are in good agreement with published numerical results and experimental data for hematocrit ratio and hematocrit and velocity profiles.

Near-Wall Migration Dynamics of Erythrocytes in Vivo: Effects of Cell Deformability and Arteriolar Bifurcation.

Red blood cell (RBC) deformability has a significant impact on microcirculation by affecting cell dynamics. Despite previous studies that have demonstrated the margination of rigid cells and particles in vitro, little information is available on the in vivo margination of deformability-impaired RBCs under physiological flow and hematocrit conditions. Thus, in this study, we examined how the deformability-dependent, RBC migration alters the cell distribution under physiological conditions, particularly in arteriolar network flows. The hardened RBCs (hRBCs) were found to preferentially flow near the vessel walls of small arterioles (diameter = 47.1–93.3 μm). The majority of the hRBCs (63%) were marginated within the range of 0.7R-0.9R (R: radial position normalized by vessel radius), indicating that the hRBCs preferentially accumulated near the vessel walls. The laterally marginated hRBCs maintained their lateral positions near the walls while traversing downstream with attenuated radial dispersion. In addition, the immediate displacement of RBCs while traversing a bifurcation also contributes to the near-wall accumulation of hRBCs. The notable difference in the inward migration between the marginated nRBCs and hRBCs after bifurcations further supports the potential role of bifurcations in the accumulation of hRBCs near the walls.

MAGNETOHYDRODYNAMIC EFFECTS ON THE GRANULAR TEMPERATURE OF RED BLOOD CELLS IN MICROVASCULATURE

The migration of Red Blood Cells (RBCs) from the wall towards the center of a narrow vessel is the result of the Fahraeus–Lindqvist effect which contemplates the dependence of viscosity and diameter. The kinetic theory explains the formation of the near-wall cell-depleted layer introducing the granular temperature that is defined as the mean square of RBCs fluctuations. The proposed mathematical model elucidates the effect of an externally applied magnetic field on the velocity and granular temperature of RBCs in a microvasculature. The effect of the volume fraction of RBCs on the velocity and granular temperature profiles is also presented and discussed. Based on the insight of the kinetic theory, the application of a stronger static magnetic field probably leads to a restriction of the migration process of RBCs towards the center of the microvessel.

Hemodynamics in stenotic vessels of small diameter under steady state conditions: Effect of viscoelasticity and migration ofredblood cells.

In microcirculation, the non-Newtonian behavior of blood and the complexity of the microvessel network are responsible for the high flow resistance and the large reduction of the blood pressure. Red blood cell aggregation along with inward radial migration are two significant mechanisms determining the former. Yet, their impact on hemodynamics in non-straight vessels is not well understood. In this study, the steady state blood flow in stenotic rigid vessels is examined, employing a sophisticated non-homogeneous constitutive law. The effect of red blood cells migration on the hydrodynamics is quantified and the constitutive model's accuracy is evaluated. A numerical algorithm based on the two-dimensional mixed finite element method and the EVSS/SUPG technique for a stable discretization of the mass and momentum conservation equations in addition to the constitutive model is employed. The numerical simulations show that a cell-depleted layer develops along the vessel wall with an almost constant thickness for slow flow conditions. This causes the reduction of the drag force and the increase of the pressure gradient as the constriction ratio decreases. Viscoelastic effects in blood flow were found to be responsible for steeper decreases of tube and discharge hematocrits as decreasing function of constriction ratio.

Perfusion pressure and blood flow determine microvascular apparent viscosity.

What is the central question of this study? The aim was to evaluate the effect of perfusion pressure on blood flow in small arterioles. The hypothesis was that blood flow regulates the thickness of the red-cell-free layer and, therefore, blood flow determines blood apparent viscosity and local vascular resistance in vascular networks with limited myogenic or metabolic regulation of blood flow. What is the main finding and its importance? Reduced perfusion pressures lowered volumetric flow rates and increased local vascular resistance, due to increased blood apparent viscosity. Thus, the local vascular resistance of small arterioles with limited myogenic or metabolic regulation of blood flow, appeared to be determined by changes in blood rheology rather than blood vessel diameter. The study of blood flow regulation is important to understand and resolve pathological conditions. As blood is a complex non-Newtonian multiphase system, the foundations of blood rheological properties have been obtained mostly in viscometers. However, blood rheological behaviour in vivo depends on the concentration of red blood cells (RBCs), their mechanical properties and the RBC hydrodynamics, including RBC migration away from the vessel wall in shear flow. This migration promotes the formation of a RBC-depleted zone, or cell-free layer (CFL), which reduces the apparent viscosity of blood. We hypothesize that perfusion pressure determines blood apparent viscosity in microvessels, as shear rate affects axial migration of RBCs by influencing the CFL thickness. In this study, we analysed the effects of perfusion pressure on blood flow in individual arterioles within the rat cremaster muscle preparation. Perfusion pressures to this microvascular bed were controlled by occlusions of the iliac artery using a pressure cuff. Blood flow measurements were obtained from direct measurements of blood flow velocity profile, as well as determination of CFL thickness using intravital microscopy. Our results indicate that perfusion pressure determines shear rates and the CFL thickness and its variations. In addition, blood flow reduction increased local vascular resistance by augmenting blood apparent viscosity rather than vascular hindrance. In conclusion, blood rheology could act as an intrinsic mechanism to further limit blood flow to tissue with limited myogenic and metabolic responses at low perfusion pressures.