Continuous dielectrophoretic size-based particle sorting

Continuous-flow dielectrophoretic (DEP) particle separation based on size is demonstrated in a microfluidic device. Polystyrene microspheres suspended in a neutrally buoyant aqueous solution are used as model particles to study DEP induced by an array of slanted, planar, interdigitated electrodes inside of a soft-lithography microchannel. The E-field gradients from the slanted electrodes impart a net transverse force component on the particles that causes them to "ratchet" across the channel. Over the length of the device, larger particles are deflected more than smaller particles according to the balance of hydrodynamic drag and DEP forces. Consequently, a flow-focused particle suspension containing different-sized particles is fractionated as the beads flow and separate down the length of the device. The flow behavior of spherical particles is modeled, and the total transverse particle displacement in the microfluidic device predicts fourth-order size and voltage and second-order inverse flow rate dependences. The model is verified experimentally for a range of flow rates, particle sizes, and E-field strengths.     

Free flow acoustophoresis: microfluidic-based mode of particle and cell separation

A novel method, free flow acoustophoresis (FFA), capable of continuous separation of mixed particle suspensions into multiple outlet fractions is presented. Acoustic forces are utilized to separate particles based on their size and density. The method is shown to be suitable for both biological and nonbiological suspended particles. The microfluidic separation chips were fabricated using conventional microfabrication methods. Particle separation was accomplished by combining laminar flow with the axial acoustic primary radiation force in an ultrasonic standing wave field. Dissimilar suspended particles flowing through the 350-microm-wide channel were thereby laterally translated to different regions of the laminar flow profile, which was split into multiple outlets for continuous fraction collection. Using four outlets, a mixture of 2-, 5-, 8-, and 10-microm polystyrene particles was separated with between 62 and 94% of each particle size ending up in separate fractions. Using three outlets and three particle sizes (3, 7, and 10 microm) the corresponding results ranged between 76 and 96%. It was also proven possible to separate normally acoustically inseparable particle types by manipulating the density of the suspending medium with cesium chloride. The medium manipulation, in combination with FFA, was further used to enable the fractionation of red cells, platelets, and leukocytes. The results show that free flow acoustophoresis can be used to perform complex separation tasks, thereby offering an alternative to expensive and time-consuming methods currently in use.     

Anticipating Cutoff Diameters in Deterministic Lateral Displacement (DLD) Microfluidic Devices for an Optimized Particle Separation

Deterministic lateral displacement (DLD) devices enable to separate nanometer to micrometer‐sized particles around a cutoff diameter, during their transport through a microfluidic channel with slanted rows of pillars. In order to design appropriate DLD geometries for specific separation sizes, robust models are required to anticipate the value of the cutoff diameter. So far, the proposed models result in a single cutoff diameter for a given DLD geometry. This paper shows that the cutoff diameter actually varies along the DLD channel, especially in narrow pillar arrays. Experimental and numerical results reveal that the variation of the cutoff diameter is induced by boundary effects at the channel side walls, called the wall effect. The wall effect generates unexpected particle trajectories that may compromise the separation efficiency. In order to anticipate the wall effect when designing DLD devices, a predictive model is proposed in this work and has been validated experimentally. In addition to the usual geometrical parameters, a new parameter, the number of pillars in the channel cross dimension, is considered in this model to investigate its influence on the particle trajectories.     

Microfluidic‐Based Approaches in Targeted Cell/Particle Separation Based on Physical Properties: Fundamentals and Applications

Cell separation is a key step in many biomedical research areas including biotechnology, cancer research, regenerative medicine, and drug discovery. While conventional cell sorting approaches have led to high‐efficiency sorting by exploiting the cell's specific properties, microfluidics has shown great promise in cell separation by exploiting different physical principles and using different properties of the cells. In particular, label‐free cell separation techniques are highly recommended to minimize cell damage and avoid costly and labor‐intensive steps of labeling molecular signatures of cells. In general, microfluidic‐based cell sorting approaches can separate cells using “intrinsic” (e.g., fluid dynamic forces) versus “extrinsic” external forces (e.g., magnetic, electric field, etc.) and by using different properties of cells including size, density, deformability, shape, as well as electrical, magnetic, and compressibility/acoustic properties to select target cells from a heterogeneous cell population. In this work, principles and applications of the most commonly used label‐free microfluidic‐based cell separation methods are described. In particular, applications of microfluidic methods for the separation of circulating tumor cells, blood cells, immune cells, stem cells, and other biological cells are summarized. Computational approaches complementing such microfluidic methods are also explained. Finally, challenges and perspectives to further develop microfluidic‐based cell separation methods are discussed.     

Gravity-driven microfluidic particle sorting device with hydrodynamic separation amplification

This paper describes a simple microfluidic sorting system that can perform size profiling and continuous mass-dependent separation of particles through combined use of gravity (1 g) and hydrodynamic flows capable of rapidly amplifying sedimentation-based separation between particles. Operation of the device relies on two microfluidic transport processes: (i) initial hydrodynamic focusing of particles in a microchannel oriented parallel to gravity and (ii) subsequent sample separation where positional difference between particles with different mass generated by sedimentation is further amplified by hydrodynamic flows whose streamlines gradually widen out due to the geometry of a widening microchannel oriented perpendicular to gravity. The microfluidic sorting device was fabricated in poly(dimethylsiloxane), and hydrodynamic flows in microchannels were driven by gravity without using external pumps. We conducted theoretical and experimental studies on fluid dynamic characteristics of laminar flows in widening microchannels and hydrodynamic amplification of particle separation. Direct trajectory monitoring, collection, and post-analysis of separated particles were performed using polystyrene microbeads with different sizes to demonstrate rapid (<1 min) and high-purity (>99.9%) separation. Finally, we demonstrated biomedical applications of our system by isolating small-sized (diameter <6 microm) perfluorocarbon liquid droplets from polydisperse droplet emulsions, which is crucial in preparing contrast agents for safe, reliable ultrasound medical imaging, tracers for magnetic resonance imaging, or transpulmonary droplets used in ultrasound-based occlusion therapy for cancer treatment. Our method enables straightforward, rapid, real-time size monitoring and continuous separation of particles in simple stand-alone microfabricated devices without the need for bulky and complex external power sources. We believe that this system will provide a useful tool to separate colloids and particles for various analytical and preparative applications and may hold potential for separation of cells or development of diagnostic tools requiring point-of-care sample preparation or testing.     

Perfusion in microfluidic cross-flow: separation of white blood cells from whole blood and exchange of medium in a continuous flow

We describe a microfluidic technique for separation of particles and cells and a device that employs this technique to separate white blood cells (WBC) from whole human blood. The separation is performed in cross-flow in an array of microchannels with a deep main channel and large number of orthogonal, shallow side channels. As a suspension of particles advances through the main channel, a perfusion flow through the side channels gradually exchanges the medium of the suspension and washes away particles that are sufficiently small to enter the shallow side channels. The microfluidic device is tested with a suspension of polystyrene beads and is shown to efficaciously exchange the carrier medium while retaining all beads. In tests with whole human blood, the device is shown to reduce the content of red blood cells (RBC) by a factor of approximately 4000 with retention of 98% of WBCs. The ratio between WBCs and RBCs reached at an outlet of the device is 2.4 on average. The device is made of a single cast of poly(dimethylsiloxane) sealed with a cover glass and is simple to fabricate. The proposed technique of separation by perfusion in continuous cross-flow could be used to enrich rare populations of cells based on differences in size, shape, and deformability.     

Design of an optimized ECCA microchannel for particle manipulation utilizing dean flow coupled elasto-inertial method

In this paper, an Eulerian-Lagrangian simulation was conducted to achieve optimal Expanded-Contracted Cavity Arrays microchannel. First, a new code was developed to solve the viscoelastic flow field, and then the particles were solved by adding appropriate forces to the OpenFOAM Lagrangian solver. This code was then validated for both Eulerian and Lagrangian models. Subsequently, the effect of different parameters such as flow rate, distance from the inlet, cavity depth and distance, and particle size were also studied to obtain the proper geometry for particle focusing. Finally, the selected channel was integrated with a straight channel to separate 4.8 and 13 μm particles. The results of current research can be used to find a proper design of an Expanded-Contracted Cavity Arrays channel to achieve precise focusing and efficient, continuous, and sheathless particle/cell separation, which is much worthy for applications such as high-speed cytometry, cell counting, sorting, and many biological applications.     

Acoustic particle filter with adjustable effective pore size for automated sample preparation

This article presents analysis and optimization of a microfluidic particle filter that uses acoustic radiation forces to remove particles larger than a selected size by adjusting the driving conditions of the piezoelectric transducer (PZT). Operationally, the acoustic filter concentrates microparticles to the center of the microchannel, minimizing undesirable particle adsorption to the microchannel walls. Finite element models predict the complex two-dimensional acoustic radiation force field perpendicular to the flow direction in microfluidic devices. We compare these results with experimental parametric studies including variations of the PZT driving frequencies and voltages as well as various particle sizes (0.5-5.0 microm in diameter). These results provide insight into the optimal operating conditions and show the efficacy of our device as a filter with an adjustable effective pore size. We demonstrate the separation of Saccharomyces cerevisiae from MS2 bacteriophage using our acoustic device. With optimized design of our microfluidic flow system, we achieved yields of greater than 90% for the MS2 with greater than 80% removal of the S. cerevisiae in this continuous-flow sample preparation device.     

On-chip diamagnetic repulsion in continuous flow

We explore the potential of a microfluidic continuous flow particle separation system based on the repulsion of diamagnetic materials from a high magnetic field. Diamagnetic polystyrene particles in paramagnetic manganese (II) chloride solution were pumped into a microfluidic chamber and their deflection behaviour in a high magnetic field applied by a superconducting magnet was investigated. Two particle sizes (5 and 10 μm) were examined in two concentrations of MnCl₂ (6 and 10%). The larger particles were repelled to a greater extent than the smaller ones, and the effect was greatly enhanced when the particles were suspended in a higher concentration of MnCl₂. These findings indicate that the system could be viable for the separation of materials of differing size and/or diamagnetic susceptibility, and as such could be suitable for the separation and sorting of small biological species for subsequent studies.     

Modelling of filtration processes of fibrous filter media

This work presents a stochastic approach, based on Monte Carlo method, to simulate liquid filtration processes through non-woven fibrous materials. The real filter material is represented as a multilayer medium with a network of multiply connected pores. To describe the deposition and resuspension of particles on and from the filter medium, the following four mechanisms were considered: particle capture by sieving, patricle capture by fibers; particle capture by blocked pores; and particle re-entrainment. The particle capture by fibers and blocked pores, and particle re-entrainment depend on the balance between the adhesion and removal forces. The adhesion forces for particles of diameter smaller than 20 μm were determined through the concept of London-Van Der Waals forces. For particles of diameter greater than 20 μm, gravitational forces were considered. Three-dimensional random flow was assumed to stimulate the particles motion through the multilayer medium. The pressure drop across the filter medium was calculated as the sum of the pressure drop across the clean filter plus the pressure drop due to the deposited particles.A FORTRAN Program was developed to implement the filtration process model. For a wide range of typical filtration conditions, the calculated filter efficiencies predicted the experimental results with a percent difference between 0.5 and 19.3 depending on the particle size. The filter material capacities were predicted with an average discrepancy of 23.0%     

Optically Coated Mirror‐Embedded Microchannel to Measure Hydrophoretic Particle Ordering in Three Dimensions

Three‐dimensional (3D) measurement of the behavior of microfluidic particles is vital for improving their operational efficiency and characterization. In particular, it is important to measure particle motions in 3D for exact characterization of hydrophoresis, which utilizes 3D convective flows for size separation. Herein, the 3D measurement of hydrophoretic particle ordering for the exact characterization of hydrophoresis by using an optically coated mirror‐embedded microchannel is reported. The mirror, ideally at 45°, reflects the side view of the channel and enables 3D positional information to be obtained easily from two different orthogonal‐axis images. With this method, it is shown that hydrophoresis is governed by convective vortices and steric hindrance. It is also observed that hydrophoresis enables 3D particle focusing without sheath flows and accurate flow‐rate control. The mechanism of hydrophoresis is finally verified by conducting a computational simulation and comparing the simulation results with the experimental measurements. The hydrophoretic method can be straightforwardly integrated as a 3D particle‐focusing component in integrated microfluidic systems. The mirror‐embedded channel can also be readily fabricated in a single cast of polydimethylsiloxane, thus offering low‐cost, easy implementation of 3D particle measurement.     

On-chip diamagnetic repulsion in continuous flow

Abstract

We explore the potential of a microfluidic continuous flow particle separation system based on the repulsion of diamagnetic materials from a high magnetic field. Diamagnetic polystyrene particles in paramagnetic manganese (II) chloride solution were pumped into a microfluidic chamber and their deflection behaviour in a high magnetic field applied by a superconducting magnet was investigated. Two particle sizes (5 and 10 μm) were examined in two concentrations of MnCl₂ (6 and 10%). The larger particles were repelled to a greater extent than the smaller ones, and the effect was greatly enhanced when the particles were suspended in a higher concentration of MnCl₂. These findings indicate that the system could be viable for the separation of materials of differing size and/or diamagnetic susceptibility, and as such could be suitable for the separation and sorting of small biological species for subsequent studies.     

Microfluidic particle sorter employing flow splitting and recombining

This paper describes an improved microfluidic device that enables hydrodynamic particle concentration and size-dependent separation to be carried out in a continuous manner. In our previous study, a method for hydrodynamic filtration and sorting of particles was proposed using a microchannel having multiple branch points and side channels, and it was applied for continuous concentration and separation of polymer particles and cells. In the current study, the efficiency of particle sorting was dramatically improved by geometrically splitting fluid flow from a main stream and recombining. With these operations, particles with diameters larger than a specific value move toward one sidewall in the mainstream. This control of particle positions is followed by the perfect particle alignment onto the sidewall, which increases the selectivity and recovery rates without using a liquid that does not contain particles. In this study, a microchannel having one inlet and five outlets was designed and fabricated. By simply introducing particle suspension into the device, concentrations of 2.1-3.0-microm particles were increased 60-80-fold, and they were collected independently from each outlet. In addition, it was demonstrated that the measured flow rates distributed into each side channel corresponded well to the theoretical values when regarding the microchannel network as a resistive circuit.     

Advection Flows‐Enhanced Magnetic Separation for High‐Throughput Bacteria Separation from Undiluted Whole Blood

A major challenge to scale up a microfluidic magnetic separator for extracorporeal blood cleansing applications is to overcome low magnetic drag velocity caused by viscous blood components interfering with magnetophoresis. Therefore, there is an unmet need to develop an effective method to position magnetic particles to the area of augmented magnetic flux density gradients while retaining clinically applicable throughput. Here, a magnetophoretic cell separation device, integrated with slanted ridge‐arrays in a microfluidic channel, is reported. The slanted ridges patterned in the microfluidic channels generate spiral flows along the microfluidic channel. The cells bound with magnetic particles follow trajectories of the spiral streamlines and are repeatedly transferred in a transverse direction toward the area adjacent to a ferromagnetic nickel structure, where they are exposed to a highly augmented magnetic force of 7.68 µN that is much greater than the force (0.35 pN) at the side of the channel furthest from the nickel structure. With this approach, 91.68% ± 2.18% of Escherichia coli (E. coli) bound with magnetic nanoparticles are successfully separated from undiluted whole blood at a flow rate of 0.6 mL h^?1 in a single microfluidic channel, whereas only 23.98% ± 6.59% of E. coli are depleted in the conventional microfluidic device.     

Toward label-free optical fractionation of blood--optical force measurements of blood cells

There is a compelling need to develop systems capable of processing blood and other particle streams for detection of pathogens that are sensitive, selective, automated, and cost/size effective. Our research seeks to develop laser-based separations that do not rely on prior knowledge, antibodies, or fluorescent molecules for pathogen detection. Rather, we aim to harness inherent differences in optical pressure, which arise from variations in particle size, shape, refractive index, or morphology, as a means of separating and characterizing particles. Our method for measuring optical pressure involves focusing a laser into a fluid flowing opposite to the direction of laser propagation. As microscopic particles in the flow path encounter the beam, they are trapped axially along the beam and are pushed upstream from the laser focal point to rest at a point where the optical and fluid forces on the particle balance. On the basis of the flow rate at which this balance occurs, the optical pressure felt by the particle can be calculated. As a first step in the development of a label-free device for processing blood, a system has been developed to measure optical pressure differences between the components of human blood, including erythrocytes, monocytes, granulocytes, and lymphocytes. Force differentials have been measured between various components, indicating the potential for laser-based separation of blood components based upon differences in optical pressure. Potential future applications include the early detection of blood-borne pathogens for the prevention of sepsis and other diseases as well as the detection of biological threat agents.     

Microfluidic characterization and continuous separation of cells and particles using conducting poly(dimethyl siloxane) electrode induced alternating current-dielectrophoresis

This paper presents a poly(dimethyl siloxane) (PDMS) polymer microfluidic device using alternating current (ac) dielectrophoresis (DEP) for separating live cells from interfering particles of similar sizes by their polarizabilities under continuous flow and for characterizing DEP behaviors of cells in stagnant flow. The ac-DEP force is generated by three-dimensional (3D) conducting PDMS composite electrodes fabricated on a sidewall of the device main channel. Such 3D PDMS composite electrodes are made by dispersing microsized silver (Ag) fillers into PDMS gel. The sidewall AgPDMS electrodes can generate a 3D electric field that uniformly distributes throughout the channel height and varies along the channel lateral direction, thereby producing stronger lateral DEP effects over the entire channel. This allows not only easy observation of cell/particle lateral motion but also using the lateral DEP force for manipulation of cells/particles. The former feature is used to characterize the frequency-dependent DEP behaviors of Saccharomyces cerevisiae (yeast) and Escherichia coli (bacteria). The latter is utilized for continuous separation of live yeast and bacterial cells from similar-size latex particles as well as live yeast cells from dead yeast cells. The separation efficiency of 97% is achieved in all cases. The demonstration of these functions shows promising applications of the microfluidic device.     

Electrokinetic measurements of particles in lubricating oils: Implications for filtrations

Experimental evidence showing the importance of eletrical double-layer forces in filtration of lubricating oil is presented. These forces are shown to be present in a wide variety of lubricating oils and are seen to be a function of the particle and fiber size, the zeta potential of the particle and fiber media, and their separation distance. The implications of these forces and their interaction with the London—van der Waals forces and the resultant interaction potential and its possible effects on the performance of oil filters is discussed.     

Particle Separation in an Annular Converging Channel with an Inner Rotating Permeable Baffle

The relation between the separation efficiency of solid particles and the stability of the helical flow of a viscous fluid in a converging channel with an inner rotating permeable cylindrical baffle has been studied. The profiles of the axial and tangential velocities and the separation efficiency of solid particles have been calculated based on the numerical solution of a system of equations describing the hydrodynamics of two-phase media. Analysis of the obtained solutions shows that vortices having an effect on particle separation can appear in the converging channel. Moreover, the larger the size of the converging annular channel, the earlier a loss of stability occurs. It has been found that the formation of vortices is impossible for some flow regimes and, as a result of fluid flow stabilization, the fraction of particles settled on the permeable cylindrical baffle decreases. It has been shown that those regime parameters at which a helical flow exists should be selected for the development of combined action units involving filtering and the separation of the solid dispersed phase.     

The Synthesis and Assembly of Polymeric Microparticles Using Microfluidics

The controlled synthesis of micrometer‐sized polymeric particles bearing features such as nonspherical shapes and spatially segregated chemical properties is becoming increasingly important. Such particles can enable fundamental studies on self‐assembly and suspension rheology, as well as be used in applications ranging from medical diagnostics to photonic devices. Microfluidics has recently emerged as a very promising route to the synthesis of such polymeric particles, providing fine control over particle shape, size, chemical anisotropy, porosity, and core/shell structure. This progress report summarizes microfluidic approaches to particle synthesis using both droplet‐ and flow‐lithography‐based methods, as well as particle assembly in microfluidic devices. The particles formed are classified according to their morphology, chemical anisotropy, and internal structure, and relevant examples are provided to illustrate each of these approaches. Emerging applications of the complex particles formed using these techniques and the outlook for such processes are discussed.     

Effect of particle size on the performance of cross-flow microfiltration

The effects of particle size on the cake properties and the performance of cross-flow microfiltration are studied. A particulate sample with a wide size distribution range from submicron to micron is used in experiments. The probabilities of particle deposition are analyzed based on a force analysis. Since themajor forces in determining the particle deposition and packing in the filter cake are different for submicron and micron particles, the particle size plays an important role in the filtration performance. Cake properties, such as mass, porosity and average specific filtration resistance of the cake, are calculated theoretically and compared with experimental data. Except for the overestimation of the mean particle size for about 1 μm, the calculated results of the pseudo-steady filtration rate and cake properties under various operating conditions agree fairly well with the experimental data.