Deterministic Lateral Displacement Research Articles

A unified-field monolithic fictitious domain-finite element method for fluid-structure-contact interactions and applications to deterministic lateral displacement problems

Based upon two overlapped, body-unfitted meshes, a type of unified-field monolithic fictitious domain-finite element method (UFMFD-FEM) is developed in this paper for moving interface problems of dynamic fluid-structure interactions (FSI), which is accompanied with high-contrast physical coefficients across the interface and with contacting collisions between the structure and fluidic channel wall in the immersed FSI case. In particular, the proposed novel numerical method consists of a monolithic, stabilized mixed finite element method within the frame of fictitious domain/immersed boundary method (FD/IBM) for generic fluid-structure-contact interaction (FSCI) problems in the Eulerian–updated Lagrangian description, while involving the no-slip type of interface conditions on the fluid-structure interface, and the repulsive contact force/stress on the structural surface when the immersed structure contacts the fluidic channel wall. The developed UFMFD-FEM for FSI/FSCI problems can deal with the structural motion with large rotational and translational displacements and/or large deformation in an accurate and efficient fashion, which is first validated by two benchmark FSI problems and one FSCI model problem, then by experimental results of a realistic FSCI scenario – the microfluidic deterministic lateral displacement (DLD) problem that is applied to isolate circulating tumor cells (CTCs) from blood cells in the blood fluid through a cascaded filter DLD microchip in practice, where a particulate fluid with the pillar obstacles effect in the fluidic channel, i.e., the effects of fluid-structure interaction and structure collision, play significant roles to sort particles (cells) of different sizes with tilted pillar arrays. The developed unified-field, monolithic fictitious domain-based mixed finite element method can be seamlessly extended to more sophisticated, high dimensional FSCI problems with contacting collisions between the moving elastic structure and fluidic channel wall.

Trajectory analysis of circulating tumor cells through contorted deterministic lateral displacement array for unruptured trapping: a simulation study

During the multistage process of cancer spread, fragments of cells branch out from the primitive tumor tissue and scatter throughout the body by blood flow stream or lymph, which are termed as circulating tumor cells (CTCs), regarded as an important biomarker for early cancer diagnosis. Deterministic lateral displacement (DLD) is one of the most efficient passive type particle isolation techniques which functions on asymmetrical laminar flow diversion around the placed microposts forming an array, based on the critical dimeter of the device. In this simulation study, complete trajectory analysis of CTCs and WBCs has been performed using COMSOL Multiphysics software with respect to the angular orientation of sample inlet channel. A simulated model of an asymmetric DLD array-based microfluidic device with latest empirical expression for critical diameter has been presented with optimized sample inlet orientation so as to maintain perfect separation efficiency of CTCs along with arrangement for unruptured trapping of CTCs and WBCs. The approximate CTC isolation efficiency was obtained above 95% for sample inlet inclinations up to 0.5°, and further tilt showed reduction in efficiency. Migration angle of CTC trajectory for the modeled device was obtained around 15° for both the sample inlet orientations leading to least intermingling of CTCs and WBCs. A smaller number of CTC–WBC interaction points was observed for clockwise sample inlet tilt compared to the anticlockwise displacement reducing possibilities of cell rupture. The shear rate around central microposts was found to be higher as well in clockwise sample inlet orientation. This simulation study establishes that sample inlet clockwise tilting of maximum around 0.5°–0.7° leads to better CTC isolation and provides better chances of unruptured trapping of cells at the designated outlets. The results of this study provide an approach toward further optimization of DLD devices' functioning and, thus, could help fabricate better DLD-based microfluidic devices for efficient trapping of CTCs.

Sheathless and high-throughput separation of multi-target particles combining inertia and deterministic lateral displacement (DLD) in a microchannel

Precisely separation of multicomponent particle mixtures has great potential to clinical diagnosis. Herein, we propose a novel two-stage separation platform combining deterministic lateral displacement (DLD) with inertia which can achieve high-throughput, sheathless, and precise multi-target particle separation. The first-stage inertial unit, consisting of contraction–expansion arrays, was used for both separating the large particles and prefocusing the remaining particles. The second-stage DLD unit was then used to further separate medium and small particles. In particular, the inertial unit can also replace sheath flow in the DLD unit to prefocus and limit the entry position of the particle flow. Moreover, flow matching of the two-stage connection region has ensured that the input flow of the second stage meets the conditions for effective separation of the DLD unit. The structure parameters and input conditions of each unit were optimized based on the evaluation of separation performance by numerical simulation and particle experiment. The results show that continuous and rapid separation of 5 μm, 10 μm and 20 μm particles can be achieved with high efficiency (96.3 %, 94.7 % and 100 %) and high purity (98.65 %, 92.65 % and 85.5 %) based on the superposition of inertia and DLD units. Our two-stage separation platform is capable of high throughput operation while maintaining separation accuracy, which has strong practical significance in biological, clinical and point-of-care applications.

A Novel Microfluidic Strategy for Efficient Exosome Separation via Thermally Oxidized Non-Uniform Deterministic Lateral Displacement (DLD) Arrays and Dielectrophoresis (DEP) Synergy.

Exosomes, with diameters ranging from 30 to 150 nm, are saucer-shaped extracellular vesicles (EVs) secreted by various type of human cells. They are present in virtually all bodily fluids. Owing to their abundant nucleic acid and protein content, exosomes have emerged as promising biomarkers for noninvasive molecular diagnostics. However, the need for exosome separation purification presents tremendous technical challenges due to their minuscule size. In recent years, microfluidic technology has garnered substantial interest as a promising alternative capable of excellent separation performance, reduced reagent consumption, and lower overall device and operation costs. In this context, we hereby propose a novel microfluidic strategy based on thermally oxidized deterministic lateral displacement (DLD) arrays with tapered shapes to enhance separation performance. We have achieved more than 90% purity in both polystyrene nanoparticle and exosome experiments. The use of thermal oxidation also significantly reduces fabrication complexity by avoiding the use of high-precision lithography. Furthermore, in a simulation model, we attempt to integrate the use of dielectrophoresis (DEP) to overcome the size-based nature of DLD and distinguish particles that are close in size but differ in biochemical compositions (e.g., lipoproteins, exomeres, retroviruses). We believe the proposed strategy heralds a versatile and innovative platform poised to enhance exosome analysis across a spectrum of biochemical applications.

Geometry Scaling for Externally Balanced Cascade Deterministic Lateral Displacement Microfluidic Separation of Multi-Size Particles.

To non-invasively monitor personal biological and environmental samples in Internet of Things (IoT)-based wearable microfluidic sensing applications, the particle size could be key to sensing, which emphasizes the need for particle size fractionation. Deterministic lateral displacement (DLD) is a microfluidic structure that has shown great potential for the size fractionation of micro- and nano-sized particles. This paper introduces a new externally balanced multi-section cascade DLD approach with a section-scaling technique aimed at expanding the dynamic range of particle size separation. To analyze the design tradeoffs of this new approach, a robust model that also accounts for practical fabrication limits is presented, enabling designers to visualize compromises between the overall device size and the achievement of various performance goals. Furthermore, results show that a wide variety of size fractionation ranges and size separation resolutions can be achieved by cascading multiple sections of an increasingly smaller gap size and critical separation dimension. Model results based on DLD theoretical equations are first presented, followed by model results that apply the scaling restrictions associated with the second order of effects, including practical fabrication limits, the gap/pillar size ratio, and pillar shape.

Investigating the impact of pillar shape on flexible erythrocyte separation through deterministic lateral displacement

The deterministic lateral displacement (DLD) method has gradually moved beyond the separation of spherical rigid particles and has been successfully applied to the field of biological cells, especially to the separation of disc-shaped flexible erythrocytes. Different shapes of micropillars in the DLD device significantly affect the sorting performance of erythrocytes. We investigated the effect of the flow field in the gap of micropillars of different shapes (circular, triangular, I-shaped) on the behavior of erythrocyte tumbling motion. The simulation results showed that in the I-shaped pillar DLD device, compared with the circular and triangular pillars, the erythrocytes exhibited a stable and continuous flipping motion during the movement along the array offset direction, which enabled the effective separation size to be maintained at a high level. In addition, the formula for calculating the critical diameter of the I-shaped pillar DLD array for separating erythrocytes was obtained by fitting, which provides strong support for a more efficient design of the I-shaped pillar DLD device dedicated to erythrocyte sorting.

On-chip label-free sorting and enrichment of microplastic particles by using deterministic lateral displacement

A novel deterministic lateral displacement (DLD) method employing a pressure-driven flow for the continuous size-based separation of microplastic particles is presented in this paper. To induce the DLD effect, arrays of triangular posts were designed to enhance the sorting resolution and reduce the particles clogging. For the particles with a diameter larger than the critical diameter (Dc) in the DLD device, they move in bump mode with collision to microposts. While, the particles flow in zigzag mode if their sizes are below the Dc value. The DLD microfluidic chip enables simplified fabrication process and shows property of label-free and high throughput. Numerical studies were conducted to discuss the Dc values in the microchannel with horizontally symmetrical and asymmetrical posts, where Dc was found smaller in the asymmetric horizontal flow, enabling higher separation sensitivity and resolution. Experiments were conducted to demonstrate the separation of 10 μm and 15 μm polystyrene microplastic particles, and different types of polystyrene and polyethylene microplastic particles by adjusting the flow rates. In order to achieve successful separation, the flow rates between the sheath flow and the sample solution were well matched. In this way, the proposed DLD microfluidic chip with horizontally asymmetrical triangular posts shows property of label-free, high throughput, capability to analyze microplastic particle selectively and sensitively, possibility of sorting nanoplastic particles.

The slow viscous flow around a general rectangular doubly-periodic arrays of infinite slender cylinders

Abstract The slow viscous flow through a doubly-periodic array of cylinders does not have an analytical solution. However, as a reduced model for the flow within fibrous porous media and microfluidic arrays, this solution is important for many real-world systems. We asymptotically determine the flow around a general rectangular doubly-periodic array of infinite slender cylinders, extending the existing asymptotic solution for square arrays. The flow in the cell is represented by a collection of doubly-periodic, rapidly-convergent two-dimensional singularity solutions, and the boundary condition on the surface of the cylinder is solved asymptotically in powers of the cylinder radius. The asymptotic solution provides an easily computed closed-form estimate for the flow and forces as a function of the radius and the dimensions of the cell. The force is compared to results from lattice-Boltzmann simulations of low-Reynolds-number flows in the same geometry, and the accuracy of the no-slip condition on the surface of the cylinder, predicted by the asymptotic theory, is assessed. Finally, the behaviour of the flow, flux, force and effective permeability of the cell is investigated as a function of the geometric parameters. The structure of the asymptotic permeability is consistent with previous single-geometry predictions but provides a closed-form estimate for how the aspect ratio of the cell changes the leading-order behaviour. These models could be used to help understand the flows within porous systems composed of fibres and systems involving periodic arrays such as systems based on deterministic lateral displacement .

Design and fabrication of a polydimethylsiloxane device for evaluating the effect of pillar geometry and configuration in the flow separation using deterministic lateral displacement.

The advancement of microfluidics and the manufacturing of microdevices has led to a strategic change in the biomedical industry. The flow through narrow channels and the pillars are placed strategically, leading to the phenomenon of particle separation through deterministic lateral displacement (DLD). In such a phenomenon, the shape, size, location and orientation of the obstacles play an important role. For the first time, particle separation is achieved with DLD modules having high row shift angles of 25°, 30° and 35°, reducing the number of pillars. The significance of circular and triangular micropillars executing deterministic lateral displacement, oriented at different angles, has been investigated, and it is found that the triangular pillars oriented at 75° resulted in better separation compared to the other configurations. In this report, the fabrication, location, orientation of the micropillars and the selection of appropriate process parameters are detailed. The structures are fabricated on silicon wafers using the standard photolithography process followed by the deep reactive ion etching process. These dies are further used to fabricate the polydimethylsiloxane-based microfluidic chips. These fabricated devices are characterised by their size, structure and quality using 3D microscopy and scanning electron microscopy. Further, blood plasma separation is carried out using the devices fabricated in this work, and the particles at the inlet and outlets are evaluated using microscopy and a novel image processing technique, replacing the use of a hemocytometer. The path traced by the particles at different flow conditions is numerically evaluated and validated with experiments. The novel device is capable of separating blood cells from plasma with a recovery factor varying from 44% to 100%. PDMS-PDMS bonding experiments using oxygen and argon plasma have been carried out to evaluate the maximum bond strength and flow velocity in the devices. It is observed that the oxygen plasma results in a bond strength of 0.404 N mm-1, thus a high throughput of 135.34 μL s-1 is achieved using the fabricated device.

Rapid cell isolation in breastmilk in a non-clinical setting by a deterministic lateral displacement device and selective water and fat absorption.

Breastmilk is a reliable source of biomarker-containing, sloughed breast cells that have the potential to give valuable health insights to new mothers. Furthermore, known DNA-based markers for pregnancy-associated breast cancer are chemically stable and can be safely stored on a commercially available FTA® Elute Micro (EM) card, which can subsequently be mailed to a testing facility for the cost of a stamp. In theory, this archiving process can be performed by nonprofessionals in very low-resource settings as it simply requires placing a drop of breastmilk on an EM card. Although this level of convenience is paramount for new mothers, the low cell density of breastmilk complicates archiving on an EM card as such commercial products and associated protocols were designed for high-cell density physiological fluids such as blood. In this study, we present the use of a deterministic lateral displacement (DLD) device combined with porous superabsorbent polymers and hydrophobic sponges to achieve simple and low-cost cell enrichment in breastmilk. As the critical separation diameter in a DLD device is more heavily dependent on lithographically controlled pillar layout than fluid or flow properties, our use of DLD microfluidics allowed for the accommodation of both varying viscosities in human breastmilk samples and a varying pressure of actuation resulting from manual, syringe-driven operation. We demonstrate successful cell enrichment (>11×) and a corresponding increase in the DNA concentration of EM card elutions among breastmilk samples processed with our hybrid microfluidic system. As our device achieves sufficiently high cell enrichment in breastmilk samples while only requiring the user to push a syringe for 4 min with reasonable effort, we believe that it has high potential to expand EM card DNA archiving for diagnostic applications with low-cell density physiological fluids and in low-resource settings.

Dielectrophoretic enrichment of live chemo-resistant circulating-like pancreatic cancer cells from media of drug-treated adherent cultures of solid tumors.

Due to low numbers of circulating tumor cells (CTCs) in liquid biopsies, there is much interest in enrichment of alternative circulating-like mesenchymal cancer cell subpopulations from in vitro tumor cultures for utilization within molecular profiling and drug screening. Viable cancer cells that are released into the media of drug-treated adherent cancer cell cultures exhibit anoikis resistance or anchorage-independent survival away from their extracellular matrix with nutrient sources and waste sinks, which serves as a pre-requisite for metastasis. The enrichment of these cell subpopulations from tumor cultures can potentially serve as an in vitro source of circulating-like cancer cells with greater potential for scale-up in comparison with CTCs. However, these live circulating-like cancer cell subpopulations exhibit size overlaps with necrotic and apoptotic cells in the culture media, which makes it challenging to selectively enrich them, while maintaining them in their suspended state. We present optimization of a flowthrough high frequency (1 MHz) positive dielectrophoresis (pDEP) device with sequential 3D field non-uniformities that enables enrichment of the live chemo-resistant circulating cancer cell subpopulation from an in vitro culture of metastatic patient-derived pancreatic tumor cells. Central to this strategy is the utilization of single-cell impedance cytometry with gates set by supervised machine learning, to optimize the frequency for pDEP, so that live circulating cells are selected based on multiple biophysical metrics, including membrane physiology, cytoplasmic conductivity and cell size, which is not possible using deterministic lateral displacement that is solely based on cell size. Using typical drug-treated samples with low levels of live circulating cells (<3%), we present pDEP enrichment of the target subpopulation to ∼44% levels within 20 minutes, while rejecting >90% of dead cells. This strategy of utilizing single-cell impedance cytometry to guide the optimization of dielectrophoresis has implications for other complex biological samples.

Portable platform for leukocyte extraction from blood using sheath-free microfluidic DLD.

Leukocyte count is routinely performed for diagnostic purposes and is rapidly emerging as a significant biomarker for a wide array of diseases. Additionally, leukocytes have demonstrated considerable promise in novel cell-based immunotherapies. However, the direct retrieval of leukocytes from whole blood is a significant challenge due to their low abundance compared to erythrocytes. Here, we introduce a microfluidic-based platform that isolates and recovers leukocytes from diluted whole blood in a single step. Our platform utilizes a novel, sheathless method to initially sediment and focus blood cells into a dense stream while flowing through a tubing before entering the microfluidic device. A hexagonal-shaped structure, patterned at the device's inlet, directs all the blood cells against the channel's outer walls. The focused cells are then separated based on their size using the deterministic lateral displacement (DLD) microfluidic technique. We evaluated various parameters that could influence leukocyte separation, including different focusing structures (assessed both computationally and experimentally), the orientation of the tubing-chip interface, the effects of blood sample hematocrit (dilution), and flow rate. Our device demonstrated the ability to isolate leukocytes from diluted blood with a separation efficiency of 100%, a recovery rate of 76%, and a purity of 80%, while maintaining a cell viability of 98%. The device operates for over 30 min at a flow rate of 2 μL min-1. Furthermore, we developed a handheld pressure controller to drive fluid flow, enhancing the operability of our platform outside of central laboratories and enabling near-patient testing. Our platform can be integrated with downstream cell-based assays and analytical methods that require high leukocyte purity (80%), ranging from cell counting to diagnostics and cell culture applications.

MarrowCellDLD: a microfluidic method for label-free retrieval of fragile bone marrow-derived cells

Functional bone marrow studies have focused primarily on hematopoietic progenitors, leaving limited knowledge about other fragile populations, such as bone marrow adipocytes (BMAds) and megakaryocytes. The isolation of these cells is challenging due to rupture susceptibility and large size. We introduce here a label-free cytometry microsystem, MarrowCellDLD, based on deterministic lateral displacement. MarrowCellDLD enables the isolation of large, fragile BM-derived cells based on intrinsic size properties while preserving their viability and functionality. Bone marrow adipocytes, obtained from mouse and human stromal line differentiation, as well as megakaryocytes, from primary human CD34+ hematopoietic stem and progenitor cells, were used for validation. Precise micrometer-range separation cutoffs were adapted for each cell type. Cells were sorted directly in culture media, without pre-labeling steps, and with real-time imaging for quality control. At least 106 cells were retrieved intact per sorting round. Our method outperformed two FACS instruments in purity and yield, particularly for large cell size fractions. MarrowCellDLD represents a non-destructive sorting tool for large, fragile BM-derived cells, facilitating the separation of pure populations of BMAds and megakaryocytes to further investigate their physiological and pathological roles.

Unraveling the motion and deformation characteristics of red blood cells in a deterministic lateral displacement device

Deterministic Lateral Displacement (DLD) device has gained widespread recognition and trusted for filtering blood cells. However, there remains a crucial need to explore the complex interplay between deformable cells and flow within the DLD device to improve its design. This paper presents an approach utilizing a mesoscopic cell-level numerical model based on dissipative particle dynamics to effectively capture this complex phenomenon. To establish the model’s credibility, a series of numerical simulations were conducted and the numerical results were validated with nominal experimental data from the literature. These include single cell stretching experiment, comparisons of the morphological characteristics of cells in DLD, and comparison the specific row-shift fraction of DLD required to initiate the zigzag mode. Additionally, we investigate the effect of cell rigidity, which serves as an indicator of cell health, on average flow velocity, trajectory, and asphericity. Moreover, we extend the existing theory of predicting zigzag mode for solid spherical particles to encompass the behavior of red blood cells. To achieve this, we introduce a new concept of effective diameter and demonstrate its applicability in providing highly accurate predictions across a wide range of conditions.

Computational analysis of differently sized circulating tumor cells seclusion using cascaded deterministic lateral displacement array approach in microfluidics

Computational analysis of differently sized circulating tumor cells seclusion using cascaded deterministic lateral displacement array approach in microfluidics

Mesoscopic simulation of multi-scheme particle separation in deterministic lateral displacement devices using two-piece hybrid pillars

Pillar shape exploration in deterministic lateral displacement (DLD) technique holds great promise for developing high-performance microfluidic devices with versatile sorting schemes. A recent innovative design using filter-like micropillars was proposed to improve cell separation, but its significance might be greatly underestimated due to an inaccurate understanding of the underlying mechanism. In this study, we employ mesoscopic hydrodynamic simulations to explore the movement and separation of rigid spherical particles in DLD arrays using various two-piece hybrid (TPH) pillars, where each pillar consists of two individual pieces separated by a tunable inter-piece channel. In comparison with the conventional one-piece pillars, the back piece of TPH-pillars is found to hierarchically tailor the flow profile of the front piece on the basis of the row shift fraction and the inter-piece channel width, resulting in unique tunable multi-scheme separation at low, intermediate, and high row shift fractions, respectively. At the intermediate regime, in particular, the first flow lane that determines the critical separation size could be physically fenced out by the inter-piece channel, and a delicate coupling of hydrodynamic filtration and DLD has been revealed to induce a constant critical size in the whole regime. This work theoretically demonstrates the feasibility and significance of TPH-pillars, which may open up a new direction of the geometry design by exploiting rich multi-piece hybrid structures to expand the versatility of the DLD technique.

Stiffness influence on particle separation in polydimethylsiloxane-based deterministic lateral displacement devices

Polydimethylsiloxane (PDMS) is a popular material to rapidly manufacture microfluidic deterministic lateral displacement (DLD) devices for particle separation. However, manufacturing and operation challenges are encountered with decreasing device dimensions required to separate submicron particles. The smaller dimensions, notably, cause high hydraulic resistance, resulting in significant pressure even at relatively low throughputs. This high pressure can lead to PDMS deformation, which, in turn, influences the device performance. These effects may often be overlooked in the design and operation of devices but provide a systematic source of error and inaccuracies. This study focuses in detail on these effects and investigates pillar deformation in detail. Subsequently, we discuss a potential solution to this deformation using thermal annealing to stiffen the PDMS. We evaluate the influence of stiffness on the separation performance at elevated sample flow rates with submicron particles (0.45 and 0.97 µm diameter). An excellent separation performance at high throughput is successfully maintained in stiffer PDMS-based DLD devices, while the conventional devices showed decreased separation performance. However, the increased propensity for delamination constrains the maximal applicable throughput in stiffer devices. PDMS deformation measurements and numerical simulations are combined to derive an iterative model for calculating pressure distribution and PDMS deformation. Finally, the observed separation characteristics and encountered throughput constraints are explained with the iterative model. The results in this study underline the importance of considering pressure-induced effects for PDMS-based DLD devices, provide a potential mitigation of this effect, and introduce an approach for estimating pressure-induced deformation.

Fabrication of Embedded Microfluidic Chips with Single Micron Resolution Using Two‐Photon Lithography

AbstractTwo‐photon lithography (TPL) is an advanced high‐resolution additive manufacturing technique for objects with feature sizes between 100 nanometers to tens of micrometers and an overall footprint of up to hundreds of micrometers. With recent advances in the TPL technique, writing speeds are becoming faster, rendering the method feasible to print high‐resolution microfluidic chips with a footprint in the centimeter range within a reasonable time frame. In this work, a process flow to fabricate embedded microfluidic chips with channel diameters down to 30 µm is developed. To address the particular difficulty of washing the embedded channels free of uncured material, introduces a developing scheme based on a 3D printed chip‐to‐world‐interface to connect the chips to a pressure‐driven pump. This setup is leakage‐free up to a pressure of 6.9 bar for faster and safer development of embedded microfluidic devices. It manufactures meander chips with channel lengths up to 20 cm, droplet generator chips, and cell sorting chips based on deterministic lateral displacement with pillar diameters of 30 µm and pillar spacing of 4 µm. TPL of microfluidic chips will enable rapid manufacturing of novel designs, significantly reducing concept‐to‐chip times with high resolution in a reasonable amount of time.

Nanostructure enabled extracellular vesicles separation and detection

Extracellular vesicles (EVs) have recently attracted significant research attention owing to their important biological functions, including cell-to-cell communication. EVs are a type of membrane vesicles that are secreted into the extracellular space by most types of cells. Several biological biomolecules found in EVs, such as proteins, microRNA, and DNA, are closely related to the pathogenesis of human malignancies, making EVs valuable biomarkers for disease diagnosis, treatment, and prognosis. Therefore, EV separation and detection are prerequisites for providing important information for clinical research. Conventional separation methods suffer from low levels of purity, as well as the need for cumbersome and prolonged operations. Moreover, detection methods require trained operators and present challenges such as high operational expenses and low sensitivity and specificity. In the past decade, platforms for EV separation and detection based on nanostructures have emerged. This article reviews recent advances in nanostructure-based EV separation and detection techniques. First, nanostructures based on membranes, nanowires, nanoscale deterministic lateral displacement, and surface modification are presented. Second, high-throughput separation of EVs based on nanostructures combined with acoustic and electric fields is described. Third, techniques combining nanostructures with immunofluorescence, surface plasmon resonance, surface-enhanced Raman scattering, electrochemical detection, or piezoelectric sensors for high-precision EV analysis are summarized. Finally, the potential of nanostructures to detect individual EVs is explored, with the aim of providing insights into the further development of nanostructure-based EV separation and detection techniques.

Biomimetic models of fish gill rakers as lateral displacement arrays for particle separation.

Ram suspension-feeding fish, such as herring, use gill rakers to separate small food particles from large water volumes while swimming forward with an open mouth. The fish gill raker function was tested using 3D-printed conical models and computational fluid dynamics simulations over a range of slot aspect ratios. Our hypothesis predicting the exit of particles based on mass flow rates, dividing streamlines (i.e. stagnation streamlines) at the slots between gill rakers, and particle size was supported by the results of experiments with physical models in a recirculating flume. Particle movement in suspension-feeding fish gill raker models was consistent with the physical principles of lateral displacement arrays ('bump arrays') for microfluidic and mesofluidic separation of particles by size. Although the particles were smaller than the slots between the rakers, the particles skipped over the vortical region that was generated downstream from each raker. The particles 'bumped' on anterior raker surfaces during posterior transport. Experiments in a recirculating flume demonstrate that the shortest distance between the dividing streamline and the raker surface preceding the slot predicts the maximum radius of a particle that will exit the model by passing through the slot. This theoretical maximum radius is analogous to the critical separation radius identified with reference to the stagnation streamlines in microfluidic and mesofluidic devices that use deterministic lateral displacement and sieve-based lateral displacement. These conclusions provide new perspectives and metrics for analyzing cross-flow and cross-step filtration in fish with applications to filtration engineering.