We developed a boundary integral formulation to simulate a red blood cell (RBC) squeezing through a submicron slit under prescribed inlet and outlet pressures. The main application of this computational study is to investigate splenic filtrations of RBCs and the corresponding in vitro mimicking microfluidic devices, during which RBCs regularly pass through inter-endothelial slits with a width less than 1.0 µm. The diseased and old RBCs are damaged or destroyed in this mechanical filtration process. We first derived the boundary integral equations of a RBC immersed in a confined domain with prescribed inlet and outlet pressures. We applied a unified self-adaptive quadrature to accurately evaluate singular and nearly singular integrals, which are especially important in this fluid-structure interaction problem with strong lubrication. A multiscale model is applied to calculate forces from the RBC membrane, and it is coupled to boundary integral equations to simulate the fluid-structure interaction. After multi-step verifications and validations against analytical and experimental results, we systematically investigated the effects of pressure drop, volume-to-surface-area ratio, internal viscosity, and membrane stiffness on RBC deformation and internal stress. We found that spectrins of RBCs could be stretched by more than 2.5 times under high hydrodynamic pressure and that the bilayer tension could be more than 500 pN/μm, which might be large enough to open mechanosensitive channels but too small to rupture the bilayer. On the other hand, we found that the bilayer-cytoskeletal dissociation stress is too low to induce bilayer vesiculation.

The splenic interendothelial slits fulfill the essential function of continuously filtering red blood cells (RBCs) from the bloodstream to eliminate abnormal and aged cells. To date, the process by which 8 μ m RBCs pass through 0.3 μ m-wide slits remains enigmatic. Does the slit caliber increase during RBC passage as sometimes suggested? Here, we elucidated the mechanisms that govern the RBC retention or passage dynamics in slits by combining multiscale modeling, live imaging, and microfluidic experiments on an original device with submicron-wide physiologically calibrated slits. We observed that healthy RBCs pass through 0.28 μ m-wide rigid slits at 37 °C. To achieve this feat, they must meet two requirements. Geometrically, their surface area-to-volume ratio must be compatible with a shape in two tether-connected equal spheres. Mechanically, the cells with a low surface area-to-volume ratio (28% of RBCs in a 0.4 μ m-wide slit) must locally unfold their spectrin cytoskeleton inside the slit. In contrast, activation of the mechanosensitive PIEZO1 channel is not required. The RBC transit time through the slits follows a − 1 and − 3 power law with in-slit pressure drop and slip width, respectively. This law is similar to that of a Newtonian fluid in a two-dimensional Poiseuille flow, showing that the dynamics of RBCs is controlled by their cytoplasmic viscosity. Altogether, our results show that filtration through submicron-wide slits is possible without further slit opening. Furthermore, our approach addresses the critical need for in vitro evaluation of splenic clearance of diseased or engineered RBCs for transfusion and drug delivery.

Inertial migration of spherical particles has been investigated extensively using experiments, theory, and computational modeling. Yet, a systematic investigation of the effect of particle shape on inertial migration is still lacking. Herein, we numerically mapped the migration dynamics of a prolate particle in a straight rectangular microchannel using smoothed particle hydrodynamics at moderate Reynolds number flows. After validation, we applied our model to 2:1 and 3:1 shape aspect ratio particles at multiple confinement ratios. Their effects on the final focusing position, rotational behavior, and transitional dynamics were studied. In addition to the commonly reported tumbling motion, for the first time, we identified a new logrolling behavior of a prolate ellipsoidal particle in the confined channel. This new behavior occurs when the confinement ratio is above an approximate threshold value of K = 0.72. Our microfluidic experiments using cell aggregates with similar shape aspect ratio and confinement ratio confirmed this new predicted logrolling motion. We also found that the same particle can undergo different rotational modes, including kayaking behavior, depending on its initial cross-sectional position and orientation. Furthermore, we examined the migration speed, angular velocity, and rotation period as well as their dependence on both particle shape aspect ratio and confinement ratio. Our findings are especially relevant to the applications where particle shape and alignment are used for sorting and analysis, such as the use of barcoded particles for biochemical assays through optical reading, or the shape-based enrichment of microalgae, bacteria, and chromosomes.

Growth of the microfluidics field has triggered numerous advances in focusing and separating microparticles, with such systems rapidly finding applications in biomedical, chemical, and environmental fields. The use of shear-thinning viscoelastic fluids in microfluidic channels is leading to evolution of elasto-inertial focusing. Herein, we showed that the interplay between the elastic and shear-gradient lift forces, as well as the secondary flow transversal drag force that is caused by the non-zero second normal stress difference, lead to different particle focusing patterns in the elasto-inertial regime. Experiments and 3D simulations were performed to study the effects of flowrate, particle size, and the shear-thinning extent of the fluid on the focusing patterns. The Giesekus constitutive equation was used in the simulations to capture the shear-thinning and viscoelastic behaviors of the solution used in the experiments. At low flowrate, with Weissenberg number Wi ~ O(1), both the elastic force and secondary flow effects push particles towards the channel center. However, at a high flowrate, Wi ~ O(10), the elastic force direction is reversed in the central regions. This remarkable behavior of the elastic force, combined with the enhanced shear-gradient lift at the high flowrate, pushes particles away from the channel center. Additionally, a precise prediction of the focusing position can only be made when the shear-thinning extent of the fluid is correctly estimated in the modeling. The shear-thinning also gives rise to the unique behavior of the inertial forces near the channel walls which is linked with the ‘warped’ velocity profile in such fluids.

The wide adoption of inertial microfluidics in biomedical research and clinical settings, such as rare cell isolation, has prompted the inquiry of its underlying mechanism. Although tremendous improvement has been made, the mechanism of inertial migration remains to be further elucidated. Contradicting observations are not fully reconciled by the existing theory, and details of the inertial migration within channel cross sections are missing in the literature. In this work, for the first time, we mapped the inertial migration pathways within channel cross section using high-speed imaging at the single-particle level. This is in contrast to the conventional method of particle streak velocimetry (PSV), which provides collective information. We also applied smoothed particle hydrodynamics (SPH) to simulate the transient motion of particles in 3D and obtained cross-sectional migration trajectories that are in agreement with the high-speed imaging results. We found two opposing pathways that explain the contradicting observations in rectangular microchannels, and the force analysis of these pathways revealed two metastable positions near the short walls that can transition into stable positions depending on the flow condition and particle size. These new findings significantly improve our understanding of the inertial migration physics, and enhance our ability to precisely control particle and cell behaviors within microchannels for a broad range of applications.

We developed coarse-grained models of spike proteins in SARS-CoV-2 coronavirus and angiotensin-converting enzyme 2 (ACE2) receptor proteins to study the endocytosis of a whole coronavirus under physiologically relevant spatial and temporal scales. We first conducted all-atom explicit-solvent molecular dynamics simulations of the recently characterized structures of spike and ACE2 proteins. We then established coarse-grained models using the shape-based coarse-graining approach based on the protein crystal structures and extracted the force field parameters from the all-atom simulation trajectories. To further analyze the coarse-grained models, we carried out normal mode analysis of the coarse-grained models to refine the force field parameters by matching the fluctuations of the internal coordinates with the original all-atom simulations. Finally, we demonstrated the capability of these coarse-grained models by simulating the endocytosis of a whole coronavirus through the host cell membrane. We embedded the coarse-grained models of spikes on the surface of the virus envelope and anchored ACE2 receptors on the host cell membrane, which is modeled using a one-particle-thick lipid bilayer model. The coarse-grained simulations show the spike proteins adopt bent configurations due to their unique flexibility during their interaction with the ACE2 receptors, which makes it easier for them to attach to the host cell membrane than rigid spikes.