Two-component macrophage model for active phagocytosis with pseudopod formation.

Macrophage phagocytosis is critical for the immune response, homeostasis regulation, and tissue repair. This intricate process involves complex changes in cell morphology, cytoskeletal reorganization, and various receptor-ligand interactions controlled by mechanical constraints. However, there is a lack of comprehensive theoretical and computational models that investigate the mechanical process of phagocytosis in the context of cytoskeletal rearrangement. To address this issue, we propose a novel coarse-grained mesoscopic model that integrates a fluid-like cell membrane and a cytoskeletal network to study the dynamic phagocytosis process. The growth of actin filaments results in the formation of long and thin pseudopods, and the initial cytoskeleton can be disassembled upon target entry and reconstructed after phagocytosis. Through dynamic changes in the cytoskeleton, our macrophage model achieves active phagocytosis by forming a phagocytic cup utilizing pseudopods in two distinct ways. We have developed a new algorithm for modifying membrane area to prevent membrane rupture and ensure sufficient surface area during phagocytosis. In addition, the bending modulus, shear stiffness, and cortical tension of the macrophage model are investigated through computation of the axial force for the tubular structure and micropipette aspiration. With this model, we simulate active phagocytosis at the cytoskeletal level and investigate the mechanical process during the dynamic interplay between macrophage and target particles.

Generation of Multiple Concentration Gradients Using a Two-Dimensional Pyramid Array.

Concentration heterogeneity of diffusible reactants is a prevalent phenomenon in biochemical processes, requiring the generation of concentration gradients for the relevant experiments. In this study, we present a high-density pyramid array microfluidic network for the effective and precise generation of multiple concentration gradients. The complex gradient distribution in the 2D array can be adaptively adjusted by modulating the reactant velocities and concentrations at the inlets. In addition, the unique design of each reaction chamber and mixing block in the array ensures uniform concentrations within each chamber during dynamic changes, enabling large-scale reactions with low reactant volumes. Through detailed numerical simulation of mass transport within the complex microchannel networks, the proposed method allows researchers to determine the desired number of reaction chambers within a given concentration range based on experimental requirements and to quickly obtain the operating conditions with the help of machine learning-based prediction. The effectiveness in generating a multiple concentration gradient environment was further demonstrated by concentration-dependent calcium carbonate crystallization experiments. This device provides a highly efficient mixing and adaptable concentration platform that is well suited for high-throughput and multiplexed reactions.

A combined computational and experimental investigation of the filtration function of splenic macrophages in sickle cell disease.

Being the largest lymphatic organ in the body, the spleen also constantly controls the quality of red blood cells (RBCs) in circulation through its two major filtration components, namely interendothelial slits (IES) and red pulp macrophages. In contrast to the extensive studies in understanding the filtration function of IES, fewer works investigate how the splenic macrophages retain the aged and diseased RBCs, i.e., RBCs in sickle cell disease (SCD). Herein, we perform a computational study informed by companion experiments to quantify the dynamics of RBCs captured and retained by the macrophages. We first calibrate the parameters in the computational model based on microfluidic experimental measurements for sickle RBCs under normoxia and hypoxia, as those parameters are not available in the literature. Next, we quantify the impact of key factors expected to dictate the RBC retention by the macrophages in the spleen, namely, blood flow conditions, RBC aggregation, hematocrit, RBC morphology, and oxygen levels. Our simulation results show that hypoxic conditions could enhance the adhesion between the sickle RBCs and macrophages. This, in turn, increases the retention of RBCs by as much as four-fold, which could be a possible cause of RBC congestion in the spleen of patients with SCD. Our study on the impact of RBC aggregation illustrates a 'clustering effect', where multiple RBCs in one aggregate can make contact and adhere to the macrophages, leading to a higher retention rate than that resulting from RBC-macrophage pair interactions. Our simulations of sickle RBCs flowing past macrophages for a range of blood flow velocities indicate that the increased blood velocity could quickly attenuate the function of the red pulp macrophages on detaining aged or diseased RBCs, thereby providing a possible rationale for the slow blood flow in the open circulation of the spleen. Furthermore, we quantify the impact of RBC morphology on their tendency to be retained by the macrophages. We find that the sickle and granular-shaped RBCs are more likely to be filtered by macrophages in the spleen. This finding is consistent with the observation of low percentages of these two forms of sickle RBCs in the blood smear of SCD patients. Taken together, our experimental and simulation results aid in our quantitative understanding of the function of splenic macrophages in retaining the diseased RBCs and provide an opportunity to combine such knowledge with the current knowledge of the interaction between IES and traversing RBCs to apprehend the complete filtration function of the spleen in SCD.

In silico and in vitro study of the adhesion dynamics of erythrophagocytosis in sickle cell disease.

Erythrophagocytosis occurring in the spleen is a critical process for removing senescent and diseased red blood cells (RBCs) from the microcirculation. Although some progress has been made in understanding how the biological signaling pathways mediate the phagocytic processes, the role of the biophysical interaction between RBCs and macrophages, particularly under pathological conditions such as sickle cell disease, has not been adequately studied. Here, we combine computational simulations with microfluidic experiments to quantify RBC-macrophage adhesion dynamics under flow conditions comparable to those in the red pulp of the spleen. We also investigate the RBC-macrophage interaction under normoxic and hypoxic conditions. First, we calibrate key model parameters in the adhesion model using microfluidic experiments for normal and sickle RBCs under normoxia and hypoxia. We then study the adhesion dynamics between the RBC and the macrophage. Our simulation illustrates three typical adhesion states, each characterized by a distinct dynamic motion of the RBCs, namely firm adhesion, flipping adhesion, and no adhesion (either due to no contact with macrophages or detachment from the macrophages). We also track the number of bonds formed when RBCs and macrophages are in contact, as well as the contact area between the two interacting cells, providing mechanistic explanations for the three adhesion states observed in the simulations and microfluidic experiments. Furthermore, we quantify, for the first time to our knowledge, the adhesive forces between RBCs (normal and sickle) and macrophages under different oxygenated conditions. Our results show that the adhesive forces between normal cells and macrophages under normoxia are in the range of 33-58 pN and 53-92 pN for sickle cells under normoxia and 155-170 pN for sickle cells under hypoxia. Taken together, our microfluidic and simulation results improve our understanding of the biophysical interaction between RBCs and macrophages in sickle cell disease and provide a solid foundation for investigating the filtration function of the splenic macrophages under physiological and pathological conditions.

In silico modeling of patient-specific blood rheology in type 2 diabetes mellitus.

Increased blood viscosity in type 2 diabetes mellitus (T2DM) is a risk factor for the development of insulin resistance and diabetes-related vascular complications; however, individuals with T2DM exhibit heterogeneous hemorheological properties, including cell deformation and aggregation. Using a multiscale red blood cell (RBC) model with key parameters derived from patient-specific data, we present a computational study of the rheological properties of blood from individual patients with T2DM. Specifically, one key model parameter, which determines the shear stiffness of the RBC membrane (µ) is informed by the high-shear-rate blood viscosity of patients with T2DM. At the same time, the other, which contributes to the strength of the RBC aggregation interaction (D0), is derived from the low-shear-rate blood viscosity of patients with T2DM. The T2DM RBC suspensions are simulated at different shear rates, and the predicted blood viscosity is compared with clinical laboratory-measured data. The results show that the blood viscosity obtained from clinical laboratories and computational simulations are in agreement at both low and high shear rates. These quantitative simulation results demonstrate that the patient-specific model has truly learned the rheological behavior of T2DM blood by unifying the mechanical and aggregation factors of the RBCs, which provides an effective way to extract quantitative predictions of the rheological properties of the blood of individual patients with T2DM.

Highly-time-resolved FMCW LiDAR with synchronously-nonlinearity-corrected acquisition for dynamic locomotion.

Highly-time-resolved and precise tracking of position, velocity, and acceleration is urgently required when highly dynamic legged robots are walking, trotting, and jumping. Frequency-modulated continuous-wave (FMCW) laser ranging is able to provide precise measurement in short distance. However, FMCW light detection and ranging (LiDAR) suffers from a low acquisition rate and poor linearity of laser frequency modulation in wide bandwidth. A sub-millisecond-scale acquisition rate and nonlinearity correction in the wide frequency modulation bandwidth have not been reported in previous studies. This study presents the synchronous nonlinearity correction for a highly-time-resolved FMCW LiDAR. The acquisition rate of 20 kHz is obtained by synchronizing the measurement signal and the modulation signal of laser injection current with a symmetrical triangular waveform. The linearization of laser frequency modulation is conducted by resampling of 1000 intervals interpolated in every up-sweep and down-sweep of 25 µs, while measurement signal is stretched or compressed in every period of 50 µs. The acquisition rate is demonstrated to be equal to the repetition frequency of laser injection current for the first time to the best of authors' knowledge. This LiDAR is successfully used to track the foot trajectory of a jumping single-leg robot. The high velocity up to 7.15 m/s and high acceleration of 365 m/s2 are measured during the up-jumping phase, while heavy shock takes place with high acceleration of 302 m/s2 as the foot end strikes the ground. The measured foot acceleration of over 300 m/s2, which is more than 30 times gravity acceleration, is reported on a jumping single-leg robot for the first time.

Ultra-sensitive all-optical phase modulation based on a fiber optic interferometer combined with Ti3C2Tx MXene-incorporated PDMS.

This study proposes an all-optical phase modulator based on a compact fiber optic interferometer combined with Ti3C2Tx MXene-incorporated PDMS. Due to the high photothermal conversion efficiency of MXene and the high thermo-optic coefficient of PDMS, changes in pump power can be effectively converted into refractive index (RI) variations in the PDMS. Then, by employing a fiber optic interferometer with a high RI response, ultra-sensitive all-optical phase modulation can be realized. The experimental results demonstrate that the resonant wavelength shift of the modulator with MXene-incorporated PDMS is 126 times higher than that of a modulator with single MXene deposition. Also, a maximum wavelength sensitivity of 14.57 nm/mW is achieved (equivalent to a phase sensitivity of 0.33π/mW); this excellent modulation performance is a great improvement on that of a previously reported fiber-based all-optical phase modulator. Furthermore, PDMS is also employed as a packaging layer to strengthen the device structure and restrict the heat in an enclosed space, which improves the heat utilization efficiency. The proposed device shows great potential in optical communication, optical filtering, sensing, and modulation applications.

Titanium Nitride Modified Fiber Optic Interferometer for Refractive Index Sensitivity Enhancement.

As one of the most well-established biocompatible transition metal nitrides, titanium nitride (TiN) has been widely applied for fiber waveguide coupling device applications. This study proposes a TiN-modified fiber optic interferometer. Benefiting from the unique properties of TiN, including ultrathin nanolayer, high refractive index, and broad-spectrum optical absorption, the refractive index (RI) response of the interferometer is greatly enhanced, which is desired all the time in the field of biosensing. The experimental results show that the deposited TiN nanoparticles (NPs) can enhance the evanescent field excitation and modulate the effective RI difference of the interferometer, which eventually results in the RI response enhancement. Besides, after incorporating the TiN with different concentrations, the resonant wavelength and the RI responses of the interferometer are enhanced to varying degrees. Benefitting from this advantage, the sensing performances, including sensitivity and measurement range, can be flexibly adapted based on different detection requirements. Since RI response can effectively reflect the detection ability of biosensors, the proposed TiN-sensitized fiber optic interferometer can be potentially applied for high-sensitive biosensing applications.

Femtosecond laser written ultra-weak Fabry-Perot array for distributed absolute temperature profile sensing with high spatial resolution.

In this paper, high spatial-resolution distributed temperature sensing has been realized based on a femtosecond laser written ultra-weak Fabry-Perot Array (FPA). 50 identical Fabry-Perot cavities are fabricated in a 10 mm long optical fiber by femtosecond laser point-by-point written technology, and the corresponding spatial resolution is as high as 200 µm. Besides, by employing the total phase demodulation method, the optical path lengths (OPLs) in the ultra-weak FPA are successively demodulated based on the Rayleigh backscattering signal recorded by an optical frequency domain reflectometry (OFDR), and therefore the absolute temperature values instead of the relative ones can be obtained. When compared with the conventional single mode fiber-based OFDR, the proposed ultra-weak FPA presents both higher spatial resolution and lower temperature sensing uncertainty (0.25 °C) benefiting from the periodically enhanced Rayleigh backscattering. Furthermore, the experiments also confirm that the ultra-weak FPA can be applied for absolute temperature field profile sensing with large temperature gradient, which is particularly suitable for high-resolution temperature measurement of miniature devices.

Ti3CN MXene-based ultra-sensitive optical fiber salinity sensor.

A Ti3CN MXene enabled ultra-sensitive optical fiber sensor is proposed, and a salinity measurement is conducted to evaluate its sensing performance in a low-concentration target molecule detection environment. Owing to the abundance of hydrophilic functional groups (-O, -F, and -OH), large specific surface, and broad-spectrum absorption characteristics of the MXene layers, the sensing performance of the MXene-incorporated sensor is greatly improved and an ultra-high salinity sensitivity of -5.34 nm/‰ is achieved (equivalent to a refractive index sensitivity of -33429 nm/RIU). Such an excellent sensing performance is 137.33% higher than that of the bare fiber sensor and is significantly enhanced over previously reported fiber sensors. Furthermore, the sensing performance of the sensor is improved without damaging the fiber structure, which is a huge advantage when compared with the traditional fiber post-processing techniques. Finally, because the refractive index is commonly used to characterize the detection ability of biosensors, our contribution suggests the integration of MXene as a potential approach to develop high-performance optical fiber biosensors.

Polycyclic Aromatic Hydrocarbons as a New Class of Promising Cathode Materials for Aluminum-Ion Batteries.

As an emerging post-lithium battery technology, aluminum ion batteries (AIBs) have the advantages of large Al reserves and high safety, and have great potential to be applied to power grid energy storage. But current graphite cathode materials are limited in charge storage capacity due to the formation of stage-4 graphite-intercalated compounds (GICs) in the fully charged state. Herein, we propose a new type of cathode materials for AIBs, namely polycyclic aromatic hydrocarbons (PAHs), which resemble graphite in terms of the large conjugated π bond, but do not form GICs in the charge process. Quantum chemistry calculations show that PAHs can bind AlCl4 - through the interaction between the conjugated π bond in the PAHs and AlCl4 - , forming on-plane interactions. The theoretical specific capacity of PAHs is negatively correlated with the number of benzene rings in the PAHs. Then, under the guidance of theoretical calculations, anthracene, a three-ring PAH, was evaluated as a cathode material for AIBs. Electrochemical measurements show that anthracene has a high specific capacity of 157 mAh g-1 (at 100 mA g-1 ) and still maintains a specific capacity of 130 mAh g-1 after 800 cycles. This work provides a feasible "theory guides practice" research model for the development of energy storage materials, and also provides a new class of promising cathode materials for AIBs.

Optimized localization algorithm of dual-Sagnac structure-based fiber optic distributed vibration sensing system.

In this study, an optimized localization algorithm is proposed for a dual-Sagnac structure-based fiber optic distributed vibration sensing (DVS) system. Different from the previous localization algorithms, the spectrum peak ratio of the interference signals in the frequency domain is applied for localization calculation, and the localization accuracy is effectively improved for the interference signal with low Signal-Noise-Ratio (SNR). Besides, the proposed optimized algorithm can solve the difficult problem of multi-point vibration localization by employing a continuous low-coherence light source, which largely reduces the system cost. Meanwhile, multi-parameter including the frequency and amplitude of the vibration signal can be retrieved simultaneously except for the vibration position, which is not available for the traditional localization algorithm of the interferometric DVS system. Experimental results verify that the system with the proposed optimized algorithm can realize high-accuracy localization of single-point vibration, multi-point with single-frequency vibration, multi-point with multi-frequency vibration. The corresponding maximum localization errors are only 0.18%, 0.22%, and 0.36% respectively.

High-sensitivity and large-range fiber optic temperature sensor based on PDMS-coated Mach-Zehnder interferometer combined with FBG.

Although numerous efforts have been dedicated towards developing fiber sensors with high performances, challenges still remain in achieving high-quality temperature sensors with high sensitivity, large measurement range and high stability. This study proposes a compact fiber optic temperature sensor based on PDMS-coated Mach-Zehnder interferometer (MZI) combined with FBG, and it can realize both high-sensitivity and large-range temperature measurement. The MZI is based on Thin No-Core Fiber (TNCF) with lateral-offset. Owing to the high refractive index sensitivity of MZI and the high thermo-optic coefficient of PDMS, the sensor can achieve a high temperature sensitivity (>10 nm/°C). Besides, by optimizing the TNCF length, the cascaded FBG can be used to locate different temperature intervals in units of approximately 10 °C, and therefore the detectable temperature range is largely extended. The experimental test demonstrates that the average sensitivities of 11.19 nm/°C, 8.53 nm/°C, 7.76 nm/°C, 7.27 nm/°C are achieved at the temperature around 30 °C, 40 °C, 50 °C and 60 °C, and it shows excellent consistency and repeatability during the thermal cycle tests.

Four-wave mixing-based photonic crystal fiber microfluid sensor with embedded U-shape microslits.

In this paper, we propose a four-wave mixing-based photonic crystal fiber (PCF) microfluid sensor, and two U-shape microslits fabricated by a femtosecond laser are embedded into the sensor for real-time microfluid measurement. Theoretical and experimental results prove that the signal wavelength is sensitive to both the refractive index (RI) and the material dispersion property of the liquid sample filled into the air channels. For different aqueous target samples at low concentrations, the responses of signal wavelength are consistent with each other. The obtained RI sensitivity is approximately 881.36 nm/RIU, and the sensing resolution is around 1.6 × 10-4 RIU. The proposed sensor also shows a better figure of merit (FOM) as high as 313.65 RIU-1 when compared with the fiber SPR sensors. Besides, the signal wavelengths present different responses with the increasing aqueous concentration due to the separated dispersion characteristics of the filled liquid samples, which can be potentially applied for the discrimination of liquid samples with a well-designed wavelength-coded sensor array in the future.

Plasmonic tapered-fiber interference sensor for simultaneously detecting refractive index and temperature.

A single-optic-fiber sensor is proposed to simultaneously detect the refractive index (RI) and temperature (T) at a single wavelength band. This sensor is based on the mixed effects of Mach-Zehnder interference (MZI) and surface plasmon resonance (SPR), where MZI is excited by a tapered-fiber structure, and SPR is stimulated by a 45 nm gold film on the tapered-fiber surface. The detection signal of an SPR spectrum superimposed on interference stripes was obtained. After fast Fourier transform and filter processing, the MZI and SPR signals were separated. Experimental results indicate that our sensor can improve the RI sensitivity to 2021.07 nm/RIU (21-fold greater than that of the original tapered-fiber MZI sensor) and detect T simultaneously. Additionally, this highly integrated sensor simplifies the detection system, with potential applications in portable biochemical sensing.

Mechanics of diseased red blood cells in human spleen and consequences for hereditary blood disorders.

In red blood cell (RBC) diseases, the spleen contributes to anemia by clearing the damaged RBCs, but its unique ability to mechanically challenge RBCs also poses the risk of inducing other pathogenic effects. We have analyzed RBCs in hereditary spherocytosis (HS) and hereditary elliptocytosis (HE), two typical examples of blood disorders that result in membrane protein defects in RBCs. We use a two-component protein-scale RBC model to simulate the traversal of the interendothelial slit (IES) in the human spleen, a stringent biomechanical challenge on healthy and diseased RBCs that cannot be directly observed in vivo. In HS, our results confirm that the RBC loses surface due to weakened cohesion between the lipid bilayer and the cytoskeleton and reveal that surface loss may result from vesiculation of the RBC as it crosses IES. In HE, traversing IES induces sustained elongation of the RBC with impaired elasticity and fragmentation in severe disease. Our simulations thus suggest that in inherited RBC disorders, the spleen not only filters out pathological RBCs but also directly contributes to RBC alterations. These results provide a mechanistic rationale for different clinical outcomes documented following splenectomy in HS patients with spectrin-deficient and ankyrin-deficient RBCs and offer insights into the pathogenic role of human spleen in RBC diseases.

Simultaneous polymerization and adhesion under hypoxia in sickle cell disease.

Polymerization and adhesion, dynamic processes that are hallmarks of sickle cell disease (SCD), have thus far been studied in vitro only separately. Here, we present quantitative results of the simultaneous and synergistic effects of adhesion and polymerization of deoxygenated sickle hemoglobin (HbS) in the human red blood cell (RBC) on the mechanisms underlying vasoocclusive pain crisis. For this purpose, we employ a specially developed hypoxic microfluidic platform, which is capable of inducing sickling and unsickling of RBCs in vitro, to test blood samples from eight patients with SCD. We supplemented these experimental results with detailed molecular-level computational simulations of cytoadherence and biorheology using dissipative particle dynamics. By recourse to image analysis techniques, we characterize sickle RBC maturation stages in the following order of the degree of adhesion susceptibility under hypoxia: sickle reticulocytes in circulation (SRs) → sickle mature erythrocytes (SMEs) → irreversibly sickled cells (ISCs). We show that (i) hypoxia significantly enhances sickle RBC adherence; (ii) HbS polymerization enhances sickle cell adherence in SRs and SMEs, but not in ISCs; (iii) SRs exhibit unique adhesion dynamics where HbS fiber projections growing outward from the cell surface create multiple sites of adhesion; and (iv) polymerization stimulates adhesion and vice versa, thereby establishing the bidirectional coupling between the two processes. These findings offer insights into possible mechanistic pathways leading to vasoocclusion crisis. They also elucidate the processes underlying the onset of occlusion that may involve circulating reticulocytes, which are more abundant in hemolytic anemias due to robust compensatory erythropoiesis.

Multiplexed Weak Waist-Enlarged Fiber Taper Curvature Sensor and Its Rapid Inline Fabrication.

This study proposes a multiplexed weak waist-enlarged fiber taper (WWFT) curvature sensor and its rapid fabrication method. Compared with other types of fiber taper, the proposed WWFT has no difference in appearance with the single mode fiber and has ultralow insertion loss. The fabrication of WWFT also does not need the repeated cleaving and splicing process, and thereby could be rapidly embedded into the inline sensing fiber without splicing point, which greatly enhances the sensor solidity. Owing to the ultralow insertion loss (as low as 0.15 dB), the WWFT-based interferometer is further used for multiplexed curvature sensing. The results show that the different curvatures can be individually detected by the multiplexed interferometers. Furthermore, it also shows that diverse responses for the curvature changes exist in two orthogonal directions, and the corresponding sensitivities are determined to be 79.1°/m-1 and -48.0°/m-1 respectively. This feature can be potentially applied for vector curvature sensing.

High-Energy Aqueous Magnesium Hybrid Full Batteries Enabled by Carrier-Hosting Potential Compensation.

Underachieved capacity and low voltage plateau is ubiquitous in conventional aqueous magnesium ion full batteries. Such limitations originate from the electrochemistry and the low carrier-hosting ((de)intercalation) potential of electrode materials. Herein, via a strategy of enhancing the electrochemistry through carrier-hosting potential compensation, high-energy Mg2+ /Na+ hybrid batteries are achieved. A Mg1.5 VCr(PO4 )3 (MVCP) cathode is coupled with FeVO4 (FVO) anode in a new aqueous/organic hybrid electrolyte, giving reliable high-voltage operation. This operation enables more sufficient (de)intercalation of hybrid carriers (Mg2+ /Na+ ), thereby enhancing the reversible capacity remarkably (233.4 mA h g-1 at 0.5 A g-1 , 92.7 Wh kg-1 electrode , that is, ≥1.75-fold higher than those in conventional aqueous electrolytes). The relatively high Na+ -hosting potential of the electrodes compensates for the low Mg2+ -hosting potential and widens/elevates the discharge plateau of the full battery up to 1.50 V. Mechanism study further reveals an unusual phase transformation of FVO to Fe2 V3 and the low-lattice-strain pseudocapacitive (de)intercalation chemistry of MVCP.

Quantifying Fibrinogen-Dependent Aggregation of Red Blood Cells in Type 2 Diabetes Mellitus.

Fibrinogen is regarded as the main glycoprotein in the aggregation of red blood cells (RBCs), a normally occurring phenomenon that has a major impact on blood rheology and hemodynamics, especially under pathological conditions, including type 2 diabetes mellitus (T2DM). In this study, we investigate the fibrinogen-dependent aggregation dynamics of T2DM RBCs through patient-specific predictive computational simulations that invoke key parameters derived from microfluidic experiments. We first calibrate our model parameters at the doublet (a rouleau consisting of two aggregated RBCs) level for healthy blood samples by matching the detaching force required to fully separate RBC doublets with measurements using atomic force microscopy and optical tweezers. Using results from companion microfluidic experiments that also provide in vitro quantitative information on cell-cell adhesive dynamics, we then quantify the rouleau dissociation dynamics at the doublet and multiplet (a rouleau consisting of three or more aggregated RBCs) levels for obese patients with or without T2DM. Specifically, we examine the rouleau breakup rate when it passes through microgates at doublet level and investigate the effect of rouleau alignment in altering its breakup pattern at multiplet level. This study seamlessly integrates in vitro experiments and simulations and consequently enhances our understanding of the complex cell-cell interaction, highlighting the importance of the aggregation and disaggregation dynamics of RBCs in patients at increased risk of microvascular complications.