Deterministic Lateral Displacement Arrays Research Articles

Label-Free Multitarget Separation of Particles and Cells under Flow Using Acoustic, Electrophoretic, and Hydrodynamic Forces.

Multiplex separation of mixed biological samples is essential in a considerable portion of biomedical research and clinical applications. An automated and operator-independent process for the separation of samples is highly sought after. There is a significant unmet need for methods that can perform fractionation of small volumes of multicomponent mixtures. Herein, we design an integrated chip that combines acoustic and electric fields to enable efficient and label-free separation of multiple different cells and particles under flow. To facilitate the connection of multiple sorting mechanisms in tandem, we investigate the electroosmosis (EO)-induced deterministic lateral displacement (DLD) separation in a combined pressure- and DC field-driven flow and exploit the combination of the bipolar electrode (BPE) focusing and surface acoustic wave (SAW) sorting modules. We successfully integrate four sequential microfluidic modules for multitarget separation within a single platform: (i) sorting particles and cells relying on the size and surface charge by adjusting the flow rate and electric field using a DLD array; (ii) alignment of cells or particles within a microfluidic channel by a bipolar electrode; (iii) separation of particles based on compressibility and density by the acoustic force; and (iv) separation of viable and nonviable cells using dielectric properties via the dielectrophoresis (DEP) force. As a proof of principle, we demonstrate the sorting of multiple cell and particle types (polystyrene (PS) particles, oil droplets, and viable and nonviable yeast cells) with high efficiency. This integrated microfluidic platform combines multiple functional components and, with its ability to noninvasively sort multiple targeted cells in a label-free manner relying on different properties, is compatible with high-definition imaging, showing great potential in diverse diagnostic and analysis applications.

Numerical simulation of critical particle size in asymmetrical deterministic lateral displacement

Microfluidics devices are widely used for particle separation. Deterministic Lateral Displacement (DLD) is a passive method for particle separation. DLD devices mainly separate particles based on their sizes. There are two main modes of movement in DLD arrays; the small particles move in a zigzag path, and the larger particles separate in the displacement mode. It is therefore important to estimate the critical particle size for the transition of modes before the fabrication of DLD devices. Asymmetry in the design of the arrays can affect the fluid behavior and the critical particle size. In this study, we investigate the effects of the asymmetry caused by changing the downstream gap size to the lateral gap size ratio on the fluid behavior and particle trajectories in DLD devices. We used two dimensional (2D) Finite Element Method (FEM) to study the variations in the flow lane's widths and combined the fluid analysis with structural mechanics to model the contact between the particles and the posts in DLD arrays. We simulated the spherical particles' trajectories with diameters ranging from 1.4 to 19.2 μm in circular post DLD arrays with a lateral gap size of 20μm. In contrast to the previous works, in these simulations, the effect of particle movement on the fluid flow profiles was considered. We evaluated the particle movement mode in seven different values of the downstream gap size to the lateral gap size ratio (ranging from 0.5 to 2) and eight different row shift fraction (ranging from 0.025 to 0.3). Our simulations showed that increasing the value of the downstream gap while the lateral gap is fixed increases the veering flow rate and width. By finding the particle with the largest diameter in the zigzag mode and the particle with the smallest diameter in the displacement mode, we estimated the critical particle diameter for each value of shift fraction in different values of the downstream gap to the lateral gap size ratio. Using these data, a curve was fitted for predicting the critical particle diameter in each ratio. Finally, a more general form of the formula for the critical particle diameter was proposed, which considers an extra parameter compared to the previous ones. The results of this study can lead to a better understanding of DLD devices' functions and, thus, save time and costs for better designs and experiments.

The automatic and high‐throughput purification and enrichment of microalgae cells using deterministic lateral displacement arrays with different post shapes

AbstractBACKGROUNDMicroalgae is an important natural resource, which has been used in various fields such as health care, chemical industry, clean energy, and environmental protection. However, microalgae samples are prone to be contaminated by bacteria, impurities, or other hybrid microalgae species. Enrichment and purification must be carried out before further research and utilization.RESULTHerein, a microfluidic microalgae enrichment and purification system based on the principle of deterministic lateral displacement (DLD) was proposed. Two different shapes of micro‐post, circular and triangular, were designed. The performance of the two chips was analyzed from many aspects, such as stability, throughput, recovery efficiency, purity, and enrichment factor. Using Platymonas cells as the samples, enrichment and purification were performed with the two chips. Experimental investigations revealed that the triangular‐shaped chip has a better performance than circular‐shaped chip; the recovery efficiency of Platymonas cells of the triangular‐shaped chip was 85.7%, the purity was 89.4%, and the maximum throughput was 500 μL/min, the enrichment factor was 92.CONCLUSIONTo our knowledge, this is the first time that DLD technology has been applied to the enrichment and purification of microalgae cells. In addition, the performance of different chips was discussed. Those findings should make a driving contribution to the enrichment and purification of microalgae, DLD technology, and the utilization of microalgae resources. © 2021 Society of Chemical Industry (SCI).

Direct Laser Writing for Deterministic Lateral Displacement of Submicron Particles

Emerging additive manufacturing (or “three-dimensional (3D) printing”) strategies offer the potential to vastly extend the capabilities of established microfluidic technologies. For example, the operational performance of “deterministic lateral displacement (DLD)” - a technique in which micro/nanoposts arrayed inside of a microfluidic channel enable passive transport of target suspended particles away from their initial flow streams - is based on geometric design variables, such as the gap spacing between the arrayed posts (G). For applications that involve DLD processing of submicron-scale particles (e.g., extracellular vesicles), however, achieving the requisite geometric control via conventional microfabrication protocols represents a technically challenging manufacturing hurdle. To bypass such barriers, here we explore the use of two-photon “direct laser writing (DLW)” for additively manufacturing DLD arrays capable of submicron particle handling. Studies of DLW fabrication conditions revealed that increasing the laser power from 22.5 mW to 27.5 mW significantly decreased G from 1.51±0.04 μm to 1.02±0.05 μm, respectively. Experimental microfluidic testing of 860 nm-in-diameter fluorescent particles within the DLW-printed DLD system revealed effective hydrodynamic railing of particles along the angled arrayed microposts, with a lateral displacement of 15.3±8.6 μm over a channel length of 500 μm. These results represent, to our knowledge, the first report of a 3D printed DLD system capable of processing submicron particles, thereby offering a promising foundation for DLW-enabled DLD-based biomedical applications.

Nanoparticle Separation through Deterministic Lateral Displacement Arrays in Poly(dimethylsiloxane)

Nanoparticle Separation through Deterministic Lateral Displacement Arrays in Poly(dimethylsiloxane)

Electrokinetic biased deterministic lateral displacement: scaling analysis and simulations

Deterministic Lateral Displacement (DLD) is a microfluidic technique where arrays of micropillars within a microchannel deflect particles leading to size-based segregation. We recently demonstrated that applying AC electric fields orthogonal to the fluid flow increases the separation capabilities of these devices with a deflection angle that depends on the electric field magnitude and frequency. Particle deviation occurs in two distinct regimes depending on frequency. At high frequencies particles deviate due to negative dielectrophoresis (DEP). At low frequencies (below 1 kHz) particles oscillate perpendicular to the flow direction due to electrophoresis and are also deflected within the device. Significantly, the threshold electric field magnitude for the low frequency deviation is much lower than for deflection at high frequencies by DEP. In order to characterize the enhanced separation at low frequencies, the induced deviation was compared between the two frequency ranges. For high frequencies, we develop both theoretically and experimentally scaling laws for the dependence of particle deviation on several parameters, namely the amplitude of the applied voltage, particle size and liquid velocity where DEP forces compete with viscous drag. A novel theoretical framework is presented that enables simulation of particle trajectories subjected to DEP forces in DLD devices. Deviation angles predicted by simulations are in very good agreement with experimental data. At low frequencies (below 1 kHz), particles follow the same scaling law, but with much lower voltages. This indicates that electrokinetic phenomena other than DEP play an important role in driving particle behaviour. Experiments show that at low frequencies, particle motion is affected by quadrupolar electrohydrodynamic flows around the insulating pillars of the DLD array. We quantify the difference between the two frequency regimes and show that an electrokinetic model based only on DEP forces is limited to frequencies of 1 kHz and above.

The fluidic resistance of an array of obstacles and a method for improving boundaries in deterministic lateral displacement arrays

Deterministic lateral displacement (DLD) is a microfluidic method of separating particles by size. DLD relies on precise flow patterns to deliver high-resolution particle separation. These patterns determine which particles are displaced laterally, and which follow the flow direction. Prior research has demonstrated that the lateral array boundaries can be designed to improve the uniformity of the critical size and hence separation performance. A DLD device with an invariant critical size throughout is yet unknown. In this work, we propose a 3D design approach. We first represent the flow through the DLD as a 2D lattice of resistors. This is used to determine the relative flow resistances at the boundaries that will deliver the correct flux patterns. We then use the lattice Boltzmann method to simulate fluid flow in a 3D unit cell of the DLD and measure the fluidic resistance for a range or typical dimensions. The results of this work are used to create a new equation for fluidic resistance as a function of post size, post height, and post spacing. We use this equation to determine array geometries that should have the appropriate resistances. We then design and simulate (in COMSOL) complete devices and measure fluid fluxes and first flow-lane widths along the boundaries. We find that the first flow-lane widths are much more uniform than in any devices described previously. This work provides the best method for designing periodic boundaries, and enables narrower, shorter, and higher throughput DLD devices.

Particle/cell separation using sheath-free deterministic lateral displacement arrays with inertially focused single straight input.

This paper proposes microfluidic particle separation by sheath-free deterministic lateral displacement (DLD) with inertial focusing in a single straight input channel. Unlike conventional DLD devices for size-based particle separation, in which sheath streams are used to focus the particles before the solution containing them reaches the DLD arrays, the proposed method uses inertial focusing to align the particles along the middle or the sidewalls of the straight rectangular input channel. The two-stage model of inertial focusing is applied to reduce the length of the side-focusing channel. The proposed method is demonstrated by using it to separate fluorescent polymer particles of diameters 13 and 7 μm, in the process of which the effect of the particle focusing regime on the separation performance is also investigated. Through middle focusing, the method is further used to separate MCF-7 cells (a model of circulating tumor cells (CTCs)) and blood cells, with ∼99.0% capture efficiency achieved.

Microfluidic Particle Sorting in Concentrated Erythrocyte Suspensions

An important step in diagnostics is the isolation of specific cells and microorganisms of interest from blood. Since such bioparticles are often present at very low concentrations, throughput needs to be as high as possible. In addition, to ensure simplicity, a minimum of sample preparation is important. Therefore, sorting schemes that function for whole blood are highly desirable. Deterministic lateral displacement (DLD) has proven to be very precise and versatile in terms of a wide range of sorting parameters. To better understand how DLD performs for blood as the hematocrit increases, we have performed measurements and simulations for spherical particles in the micrometer range moving through DLD arrays for different flow velocities and hematocrits ranging from pure buffer to whole blood. We find that the separation function of the DLD array is sustained, even though blood cells introduce a shift in the trajectories and a significant dispersion for particles that are close to the critical size in the device. Simulations qualitatively replicate our experimental observations and help us identify fundamental mechanisms for the effect of hematocrit on the performance of the DLD device.

On the design of deterministic dielectrophoresis for continuous separation of circulating tumor cells from peripheral blood cells.

Detection and analysis of circulating tumor cells (CTCs) have emerged as a promising way to diagnose cancer, study its cellular mechanism, and test or develop potential treatments. However, the rarity of CTCs among peripheral blood cells is a big challenge toward CTC detection. In addition, in cases where there is similar size range between certain types of CTCs (e.g. breast cancer cells) and white blood cells (WBCs), high-resolution techniques are needed. In the present work, we propose a deterministic dielectrophoresis (DEP) method that combines the concept of deterministic lateral displacement (DLD) and insulator-based dielectrophoresis (iDEP) techniques that rely on physical markers such as size and dielectric properties to differentiate different type of cells. The proposed deterministic DEP technology takes advantage of frequency-controlled AC electric field for continuous separation of CTCs from peripheral blood cells. Utilizing numerical modeling, different aspects of coupled DLD-DEP design such as the required applied voltages, velocities, and geometrical parameters of DLD arrays of microposts are investigated. Regarding the inevitable difference and uncertainty ranges for the reported crossover frequencies of cells, a comprehensive analysis is conducted on applied electric field frequency as design's determinant factor. Deterministic DEP design provides continuous sorting of CTCs from WBCs even with similar size and has the future potential for high throughput and efficiency.

Gel-on-a-chip: continuous, velocity-dependent DNA separation using nanoscale lateral displacement.

We studied the trajectories of polymers being advected while diffusing in a pressure driven flow along a periodic pillar nanostructure known as nanoscale deterministic lateral displacement (nanoDLD) array. We found that polymers follow different trajectories depending on their length, flow velocity and pillar array geometry, demonstrating that nanoDLD devices can be used as a continuous polymer fractionation tool. As a model system, we used double-stranded DNA (dsDNA) with various contour lengths and demonstrated that dsDNA in the range of 100-10 000 base pairs (bp) can be separated with a size-selective resolution of 200 bp. In contrast to spherical colloids, a polymer elongates by shear flow and the angle of polymer trajectories with respect to the mean flow direction decreases as the mean flow velocity increases. We developed a phenomenological model that explains the qualitative dependence of the polymer trajectories on the gap size and on the flow velocity. Using this model, we found the optimal separation conditions for dsDNA of different sizes and demonstrated the separation and extraction of dsDNA fragments with over 75% recovery and 3-fold concentration. Importantly, this velocity dependence provides a means of fine-tuning the separation efficiency and resolution, independent of the nanoDLD pillar geometry.

Integrated Microfluidic Chip for Efficient Isolation and Deformability Analysis of Circulating Tumor Cells

AbstractEfficient isolation and downstream analysis of circulating tumor cells (CTCs) beyond counting is challenging. Here, an integrated microfluidic chip for efficient isolation and deformability analysis of CTCs is reported. The microfluidic chip integrates two modules: one module is two mirrored deterministic lateral displacement (DLD) arrays for the depletion of blood cells; the other module is another DLD array with an increasing tilt angle for CTCs separation based on size and deformability. The chip allows continuous isolation of CTCs with >90% capture yield and >50% capture purity at a throughput of 12 mL h−1. Results show that the captured cancer cells maintain good viability. Downstream deformability analysis validates that invasive MDA‐MB‐231 cells are more flexible than noninvasive MCF‐7 cells. The deformability of CTCs isolated from cancer patient samples is highly heterogeneous. The implementation of cell deformability analysis integrated with microfluidics extends the laboratory capability beyond simple counting of CTCs.

Deterministic lateral displacement (DLD) in the high Reynolds number regime: high-throughput and dynamic separation characteristics

Recent progress in the development of biosensors has created a demand for high-throughput sample preparation techniques that can be easily integrated into microfluidic or lab-on-a-chip platforms. One mechanism that may satisfy this demand is deterministic lateral displacement (DLD), which uses hydrodynamic forces to separate particles based on size. Numerous medically relevant cellular organisms, such as circulating tumor cells (10–15 µm) and red blood cells (6–8 µm), can be manipulated using microscale DLD devices. In general, these often-viscous samples require some form of dilution or other treatment prior to microfluidic transport, further increasing the need for high-throughput operation to compensate for the increased sample volume. However, high-throughput DLD devices will require a high flow rate, leading to an increase in Reynolds numbers (Re) much higher than those covered by existing studies for microscale (≤ 100 µm) DLD devices. This study characterizes the separation performance for microscale DLD devices in the high-Re regime (10 < Re < 60) through numerical simulation and experimental validation. As Re increases, streamlines evolve and microvortices emerge in the wake of the pillars, resulting in a particle trajectory shift within the DLD array. This differs from previous DLD works, in that traditional models only account for streamlines that are characteristic of low-Re flow, with no consideration for the transformation of these streamlines with increasing Re. We have established a trend through numerical modeling, which agrees with our experimental findings, to serve as a guideline for microscale DLD performance in the high-Re regime. Finally, this new phenomenon could be exploited to design passive DLD devices with a dynamic separation range, controlled simply by adjusting the device flow rate.

Fluorescent label-free quantitative detection of nano-sized bioparticles using a pillar array

Disease diagnostics requires detection and quantification of nano-sized bioparticles including DNA, proteins, viruses, and exosomes. Here, a fluorescent label-free method for sensitive detection of bioparticles is explored using a pillar array with micrometer-sized features in a deterministic lateral displacement (DLD) device. The method relies on measuring changes in size and/or electrostatic charges of 1 µm polymer beads due to the capture of target bioparticles on the surface. These changes can be sensitively detected through the lateral displacement of the beads in the DLD array, wherein the lateral shifts in the output translates to a quantitative measurement of bioparticles bound to the bead. The detection of albumin protein and nano-sized polymer vesicles with a concentration as low as 10 ng mL−1 (150 pM) and 3.75 μg mL−1, respectively, is demonstrated. This label-free method holds potential for point-of-care diagnostics, as it is low-cost, fast, sensitive, and only requires a standard laboratory microscope for detection.

Dynamic control of particle separation in deterministic lateral displacement separator with viscoelastic fluids

We proposed an innovative method to achieve dynamic control of particle separation by employing viscoelastic fluids in deterministic lateral displacement (DLD) arrays. The effects of shear-thinning and elasticity of working fluids on the critical separation size in DLD arrays are investigated. It is observed that each effect can lead to the variation of the critical separation size by approximately 40%. Since the elasticity strength of the fluid is related to the shear rate, the dynamic control can for the first time be easily realized through tuning the flow rate in microchannels.

High-throughput production of satellite-free droplets through a parallelized microfluidic deterministic lateral displacement device

We present a parallel numbering-up device for microfluidic production of satellite droplet-free emulsions. The device consists of two stacked layers made of polydimethylsiloxane (PDMS). The bottom layer combines eight parallel sets of a microfluidic droplet generator (MFDG) and deterministic lateral displacement (DLD) array for size-based separation of main and satellite droplets. The top layer has input reservoirs to supply both the disperse and continuous phases evenly to the eight channels of the bottom layer. We employed 3D flow simulations to find the reservoirs most suitable for equal inflow distribution. Water-in-oil (W/O) droplets comprised of main droplets (∼67 μm in diameter) and satellite droplets (1–20 μm) were generated in the parallelized MFDGs, and they were then completely separated through the DLD arrays having a critical diameter of Dc = 37.1 μm. Thus, we confirmed production of satellite droplet-free emulsion droplets at 0.2 ml/h, eight times the processing throughput of single device (= 0.025 ml/h) without decrease in performance.

Integrated nanoscale deterministic lateral displacement arrays for separation of extracellular vesicles from clinically-relevant volumes of biological samples.

Extracellular vesicles (EVs) offer many opportunities in early-stage disease diagnosis, treatment monitoring, and precision therapy owing to their high abundance in bodily fluids, accessibility from liquid biopsy, and presence of nucleic acid and protein cargo from their cell of origin. Despite their growing promise, isolation of EVs for analysis remains a labor-intensive and time-consuming challenge given their nanoscale dimensions (30-200 nm) and low buoyant density. Here, we report a simple, size-based EV separation technology that integrates 1024 nanoscale deterministic lateral displacement (nanoDLD) arrays on a single chip capable of parallel processing sample fluids at rates of up to 900 μL h-1. Benchmarking the nanoDLD chip against commonly used EV isolation technologies, including ultracentrifugation (UC), UC plus density gradient, qEV size-exclusion chromatography (Izon Science), and the exoEasy Maxi Kit (QIAGEN), we demonstrate a superior yield of ∼50% for both serum and urine samples, representing the ability to use smaller input volumes to achieve the same number of isolated EVs, and a concentration factor enhancement of up to ∼3× for both sample types, adjustable to ∼60× for urine through judicious design. Further, RNA sequencing was carried out on nanoDLD- and UC-isolated EVs from prostate cancer (PCa) patient serum samples, resulting in a higher gene expression correlation between replicates for nanoDLD-isolated EVs with enriched miRNA, decreased rRNA, and the ability to detect previously reported RNA indicators of aggressive PCa. Taken together, these results suggest nanoDLD as a promising alternative technology for fast, reproducible, and automatable EV-isolation.

Broken flow symmetry explains the dynamics of small particles in deterministic lateral displacement arrays.

Deterministic lateral displacement (DLD) is a technique for size fractionation of particles in continuous flow that has shown great potential for biological applications. Several theoretical models have been proposed, but experimental evidence has demonstrated that a rich class of intermediate migration behavior exists, which is not predicted. We present a unified theoretical framework to infer the path of particles in the whole array on the basis of trajectories in a unit cell. This framework explains many of the unexpected particle trajectories reported and can be used to design arrays for even nanoscale particle fractionation. We performed experiments that verify these predictions and used our model to develop a condenser array that achieves full particle separation with a single fluidic input.

Maximizing particle concentration in deterministic lateral displacement arrays.

We present an improvement to deterministic lateral displacement arrays, which allows higher particle concentration enhancement. We correct and extend previous equations to a mirror-symmetric boundary. This approach allows particles to be concentrated into a central channel, no wider than the surrounding gaps, thereby maximizing the particle enrichment. The resulting flow patterns were, for the first time, experimentally measured. The performance of the device with hard micro-spheres and cells was investigated. The observed flow patterns show important differences from our model and from an ideal pattern. The 18 μm gap device showed 11-fold enrichment of 7 μm particles and nearly perfect enrichment-of more than 50-fold-for 10 μm particles and Jurkat cells. This work shows a clear path to achieve higher-than-ever particle concentration enhancement in a deterministic microfluidic separation system.

Anisotropic permeability in deterministic lateral displacement arrays.

We uncover anisotropic permeability in microfluidic deterministic lateral displacement (DLD) arrays. A DLD array can achieve high-resolution bimodal size-based separation of microparticles, including bioparticles, such as cells. For an application with a given separation size, correct device operation requires that the flow remains at a fixed angle to the obstacle array. We demonstrate via experiments and lattice-Boltzmann simulations that subtle array design features cause anisotropic permeability. Anisotropic permeability indicates the microfluidic array's intrinsic tendency to induce an undesired lateral pressure gradient. This can cause an inclined flow and therefore local changes in the critical separation size. Thus, particle trajectories can become unpredictable and the device useless for the desired separation task. Anisotropy becomes severe for arrays with unequal axial and lateral gaps between obstacle posts and highly asymmetric post shapes. Furthermore, of the two equivalent array layouts employed with the DLD, the rotated-square layout does not display intrinsic anisotropy. We therefore recommend this layout over the easier-to-implement parallelogram layout. We provide additional guidelines for avoiding adverse effects of anisotropy on the DLD.