Precisely sized separation of multiple particles based on the dielectrophoresis gradient in the z-direction

We present a new 3D dielectrophoresis-field-flow fraction (DEP-FFF) concept to achieve precise separation of multiple particles by using AC DEP force gradient in the z-direction. The interlaced electrode array was placed at the upstream of the microchannel, which not only focused the particles into a single particle stream to be at the same starting position for further separation, but also increased the spacing between each particle by the retard effect to reduce particle–particle aggregation. An inclined electrode was also designed in back of the focusing component to continuously and precisely separate different sizes of microparticles. Different magnitudes of DEP force are induced at different positions in the z-direction of the DEP gate, which causes different penetration times and positions of particles along the inclined DEP gate. 2, 3, 4, and 6?μm polystyrene beads were precisely sized fractionation to be four particle streams based on their different threshold DEP velocities that were induced by the field gradient in the z-direction when a voltage of 6.5?V_p–p was applied at a flow rate of 0.6?μl/min. Finally, Candida albicans were also sized separated to be three populations for demonstrating the feasibility of this platform in biological applications. The results showed that a high resolution sized fractionation (only 25% size difference) of multiple particles can be achieved in this DEP-based microfluidic device by applying a single AC electrical signal.     

A microfluidic device for rapid concentration of particles in continuous flow by DC dielectrophoresis

This article presents a microfluidic device (so called concentrator) for rapid and efficient concentration of micro/nanoparticles using direct current dielectrophoresis (DC DEP) in continuous fluid flow. The concentrator is composed of a series of microchannels constructed with PDMS-insulating microstructures to focus efficiently the electric field in the flow direction to provide high field strength and gradient. Multiple trapping regions are formed within the concentrator. The location of particle trapping depends on the strength of the electric field applied. Under the experimental conditions, both streaming movement and DEP trapping of particles simultaneously take place within the concentrator at different regions. The former occurs upstream and is responsible for continuous transport of the particles, whereas the latter occurs downstream and rapidly traps the particles delivered from upstream. The observation agrees with the distribution of the simulated electric field and DEP force. The performance of the device is demonstrated by successfully and effectively concentrating fluorescent nanoparticles. At the sufficiently high electric field, the device demonstrates a trapping efficiency of 100%, which means downstream DEP traps and concentrates all (100%) the incoming particles from upstream. The trapping efficiency of the device is further studied by measuring the fluorescence intensity of concentrated particles in the channel. Typically, the fluorescence intensity becomes saturated in Trap 1 by applying the voltage (400 V) for >2 min, demonstrating that rapid concentration of the nanoparticles (10⁷ particles/ml) is achieved in the device. The microfluidic concentrator described can be implemented in applications where rapid concentration of targets is needed such as concentrating cells for sample preparation and concentrating molecular biomarkers for detection.     

Bioparticle separation and manipulation using dielectrophoresis

Manipulation and separation of micro-sized particles, particularly biological particles, using the dielectrophoretic (DEP) force is an emerging technique in MEMS technology. This paper presents a DEP-based microsystem for the selective manipulation and separation of bioparticles using dielectrophoretic effects. The microfabricated DEP device consists of a sandwich structure, in which a microchannel with electrode array lining on its bottom is sandwiched between the substrate and the glass lid. Dielectrophoretic behavior of polystyrene particles with diameter of 4.3 μm was studied. Both positive DEP and negative DEP were observed. Particles under positive DEP were attracted to the edges of the electrodes, while those under negative DEP were repelled away from the electrodes and levitated at certain height above the electrodes (within a proper range of frequencies of the electric field). Levitation height of the particles was measured. It was demonstrated that the levitation height of a specific particle strongly depends on the combined contributions of a number of parameters, such as the frequency of the electric field, dielectric properties of the particles and the surrounding medium. Different particles can be separated and manipulated on the basis of their difference in these parameters.     

A cell electrofusion microfluidic chip with micro-cavity microelectrode array

A new cell electrofusion microfluidic chip with 19,000 pairs of micro-cavity structures patterned on vertical sidewalls of a serpentine-shaped microchannel has been designed and fabricated. In each micro-cavity structure, the two sidewalls perpendicular to the microchannel are made of SiO₂ insulator, and that parallel to the microchannel is made of silicon as the microelectrode. One purpose of the design with micro-cavity microelectrode array is to obtain high membrane voltage occurring at the contact point of two paired cells, where cell fusion takes place. The device was tested to electrofuse NIH3T3 and myoblast cells under a relatively low voltage (~9 V). Under an AC electric field applied between the pair of microelectrodes positioned in the opposite micro-cavities, about 85–90 % micro-cavities captured cells, and about 60 % micro-cavities are effectively capable of trapping the desired two-cell pairs. DC electric pulses of low voltage (~9 V) were subsequently applied between the micro-cavity microelectrode arrays to induce electrofusion. Due to the concentration of the local electric field near the micro-cavity structure, fusion efficiency reaches about 50 % of total cells loaded into the device. Multi-cell electrofusion and membrane rupture at the end of cell chains are eliminated through the present novel design.     

Particle streaming and separation using dielectrophoresis through discrete periodic microelectrode array

This study presents a particle manipulation and separation technique based on dielectrophoresis principle by employing an array of isosceles triangular microelectrodes on the bottom plate and a continuous electrode on the top plate. These electrodes generate non-uniform electric fields transversely across the microchannel. The particles within the flowing fluid experience a dielectrophoretic force perpendicular to the fluid flow direction due to the non-uniform electric fields. The isosceles triangular microelectrodes were designed to continuously exert a small dielectrophoretic force on the particles. Particles experiencing a larger dielectrophoretic force would move further in the perpendicular direction to the fluid flow as they traveled past each microelectrode. Polystyrene microspheres were used as the model particles, with particles of ∅20 μm employed for studying the basic characteristics of this technique. Particle separation was subsequently demonstrated on ∅10 and ∅15 μm microspheres. Using an applied sinusoidal voltage of 20 V_pp and frequency of 1 MHz, a mean separation distance of 0.765 mm between them was achieved at a flow rate of 3 μl/min (~1 mm/s), an important consideration for high throughput separation capability in a micro-scale technology device. This unique isosceles triangular microelectrodes design allows heterogeneous particle populations to be separated into multiple streams in a single continuous operation.     

Continuous size-based focusing and bifurcating microparticle streams using a negative dielectrophoretic system

Dielectrophoresis (DEP) is an electrokinetic phenomenon which is used for manipulating micro- and nanoparticles in micron-sized devices with high sensitivity. In recent years, electrode-based DEP by patterning narrow oblique electrodes in microchannels has been used for particle manipulation. In this theoretic study, a microchannel with triangular electrodes is presented and a detailed comparison with oblique electrodes is made. For each shape, the behavior of particles is compared for three different configurations of applied voltages. Electric field, resultant DEP force, and particle trajectories for configurations are computed by means of Rayan native code. The separation efficiency of the two systems is assessed and compared afterward. The results demonstrate higher lateral DEP force, responsible for particle separation, distributed wider across the channel width for triangular shape electrodes in comparison with the oblique ones. The proposed electrode shape also shows the ability of particle separation by attracting negative DEP particles to or propelling them from the flow centerline, according to the configuration of applied voltages. A major deficiency of the oblique electrodes, which is the streamwise variation of the lateral DEP force direction near the electrodes, is also eliminated in the proposed electrode shape. In addition, with a proper voltages configuration, the triangular electrodes require lower voltages for particle focusing in comparison with the oblique ones.     

Design of new microchannel to reduce applied voltage for microparticles separation using dielectrophoresis method

This paper presents a new structure of microchannel in order to reduce the applied voltage using dielectrophoresis (DEP). DEP is one of the most popular techniques to separate microparticles which needs an electric field in microfluidic devices. In this study, the AC-DEP sidewall electrodes are used. The novelty of this research is to change the outlet microchannel size which effectively reduces applied voltage. In previous work, in order to separate particles with 3.5 and 4 µm diameters, 4.5 V was needed. In new design, we keep all effective parameters constant and change one of the outlet microchannel size from 50 to 60, 70 and 80 µm. Therefore, in order to separate the microparticles, we need only 3, 2 and 1.3 V, respectively.

     

Stepwise gray-scale light-induced electric field gradient for passive and continuous separation of microparticles

This article presents a gray-scale light-induced dielectrophoresis (GS-LIDEP) method that induces the lateral displacements normal to the through-flow for continuous and passive separation of microparticles. In general, DEP force only can affect the particles within very local areas due to the electric field is exponentially decayed by the distance away from the electrodes. Unlike with conventional LIDEP, a broad-ranged electrical field gradient can easily be created by GS pattern illumination, which induces DEP forces with two directions for continuous separation of particles to their specific sub-channels. Candia albicans were effectively guided to the specific outlet with the efficiency of 90% to increase the concentration of the sample below the flow rate of 0.6?μl/min. 2 and 10?μm polystyrene particles can also be passively and well separated using the multi-step GS pattern through positive and negative DEP forces, respectively, under an applied voltage of 36?V_p–p at the frequency of 10?kHz. GS-LIDEP generated a wide-ranged DEP force that is capable of working on the entire area of the microchannel, and thus the mix of particles can be passively and continuously separated toward the opposite directions by the both positive and negative GS-LIDEP forces. This simple, low cost, and flexible separation/manipulation platform could be very promising for many applications, such as in-field detections/pretreatments.     

Sequential Field-Flow Cell Separation Method in a Dielectrophoretic Chip With 3-D Electrodes

This paper presents a sequential dielectrophoretic field-flow separation method for particle populations using a chip with a 3-D electrode structure. A unique characteristic of our chip is that the walls of the microfluidic channels also constitute the device's electrodes. This property confers the opportunity to use the electrodes' shape to generate not only the electric field gradient required for dielectrophoretic force but also a fluid velocity gradient. This interesting combination gives rise to a new solution for the dielectrophoretic separation of two particle populations. The proposed sequential field-flow separation method consists of four steps. First, the microchannel is filled with the mixture of the two populations of particle. Second, the particle populations are trapped in different locations of the microfluidic channels. The population, which exhibits positive dielectrophoresis (DEP), is trapped in the area where the distance between the electrodes is the minimum, while the other population that exhibits negative DEP is trapped in locations of maximum distance between electrodes. In the next step, increasing the flow in the microchannels will result in an increased hydrodynamic force that sweeps the cell population trapped by positive DEP out of the chip. In the last step, the electric field is removed, and the second population is swept out and collected at the outlet. For theoretical and experimental exemplification of the separation method, a population of viable and nonviable yeast cells was considered.     

Sedimentation pinched-flow fractionation for size- and density-based particle sorting in microchannels

A simple and efficient device for density-based particle sorting is in high demand for the purification of specific cells, bacterium, or environmental particles for medical, biochemical, and industrial applications. Here we present microfluidic systems to achieve size- and density-based particle separation by adopting the sedimentation effect for a size-based particle sorting technique utilizing microscale hydrodynamics, called ??pinched-flow fractionation (PFF).?? Two schemes are presented: (a) the particle inertia scheme, which utilizes the inertial force of particle movement induced by the momentum change in the curved microchannel, and (b) the device rotation scheme, in which rotation of the microdevice exerts centrifugal force on the flowing particles. In the experiments, we successfully demonstrated continuous sorting of microparticles according to size and density by using these two schemes, and showed that the observed particle movements were in good agreement with the theoretical estimations. The presented schemes could potentially become one of the functional components for integrated bioanalysis systems that can manipulate/separate small amount of precious biological samples.     

Three-dimensional analysis and enhancement of continuous magnetic separation of particles in microfluidics

In continuous magnetic separation process, particles can be deflected and separated from the direction of laminar flow by means of magnetic force depending on their magnetic susceptibility and size as well as the flow rate. To analyze and control dynamic behavior of these particles flowing in microchannels, a three-dimensional numerical model was proposed and solved for obtaining the particle trajectories under the action of a gradient magnetic field and flow field. The magnetic force distribution and particle trajectories obtained were firstly verified by analytical and experimental results. Then, a detailed analysis for the enhancement of the continuous magnetic separation efficiency by optimizing the flow parameters and microchannel configurations was carried out. The results show that the separation efficiency can be greatly improved by controlling the flow rate ratio of the two fluid streams and introducing a broadened segment in the T-shaped microchannel. And it has been demonstrated to be effective through the sorting of 2-μm and 5-μm non-magnetic particles suspended in a dilute ferrofluid by a permanent magnet. The results reported could be encouraging for the design and optimization of efficient microfluidic separation systems.     

SU-8 microchannels for live cell dielectrophoresis improvements

In this work a novel highly precise SU-8 fabrication technology is employed to construct microfluidic devices for sensitive dielectrophoretic (DEP) manipulation of budding yeast cells. A benchmark microfluidic live cell sorting system is presented, and the effect of microchannel misalignment above electrode topologies on live cell DEP is discussed in detail. Simplified model of budding Saccharomyces cerevisiae yeast cell is presented and validated experimentally in fabricated microfluidic devices. A novel fabrication process enabling rapid prototyping of microfluidic devices with well-aligned integrated electrodes is presented and the process flow is described. Identical devices were produced with standard soft-lithography processes. In comparison to standard PDMS based soft-lithography, an SU-8 layer was used to construct the microchannel walls sealed by a flat sheet of PDMS to obtain the microfluidic channels. Direct bonding of PDMS to SU-8 surface was achieved by efficient wet chemical silanization combined with oxygen plasma treatment of the contact surface. The presented fabrication process significantly improved the alignment of the microstructures. While, according to the benchmark study, the standard PDMS procedure fell well outside the range required for reasonable cell sorting efficiency. In addition, PDMS delamination above electrode topologies was significantly decreased over standard soft-lithography devices. The fabrication time and costs of the proposed methodology were found to be roughly the same.

     

集成惯性聚焦结构的介电泳微流控芯片的粒子连续分离

设计并制造了一种带有惯性聚焦结构的介电泳微流控芯片,以实现不同介电性质的粒子连续分离.采用MEMS工艺制作了介电泳微流控芯片:通道入口侧壁设置一对梯形结构使经过的粒子受惯性升力的作用聚焦到通道两侧;通道底部光刻一组夹角为90°的倾斜叉指电极产生非均匀电场,利用介电泳力和流体曳力的合力使通道两侧不同的粒子发生角度不同的偏转进入不同通道,从而实现分离.将酵母菌细胞和聚苯乙烯小球作为实验样本,分析了流速和交流电压对分离的影响,确定了二者分离的最优条件并进行分离.实验结果表明,将电导率为20μS/cm的样本溶液以5μL/min的流速注入到通道中,施加6 Vp-p、10 kHz的正弦信号,酵母菌细胞沿电极运动至夹角处后沿通道中心排出,聚苯乙烯小球沿通道两侧排出,成功实现分离,平均分离效率达92.8％、平均分离纯度达90.7％.     

Selective position of individual cells without lysis on a circular window array using dielectrophoresis in a microfluidic device

We trapped individual cells between two circular windows using negative dielectrophoretic (DEP) force and then sequentially trapped them inside circular windows by positive DEP force without electrical lysis in a microfluidic device. Three parameters, (1) the transmembrane potential that determines the lysis of a cell, (2) individual cell size that affects the trapped position accuracy of the cell, and (3) the Clausius–Mossotti (CM) factor that decides the trapped efficiency of the cell, were characterized experimentally and numerically in this sequential cell trapping technique. In this characterization, we confirmed that the swap rate of applied voltage frequency, size similarity between the cell and circular window, and instantaneous change rate of Re(f _CM) as a function of frequency were important factors in determining the selective position of individual cells without lysis. Our results provide useful suggestions for designing the structure of microfluidic DEP devices and optimizing variables required to manipulate individual cell trapping using both negative and positive DEP forces.     

Label-free continuous lateral magneto-dielectrophoretic microseparators for highly efficient enrichment of circulating nucleated cells from peripheral blood

We have developed a continuous lateral magneto-dielectrophoretic (MAP-DEP) microseparator for the highly efficient enrichment of circulating nucleated cells from peripheral blood, based on native magnetic and dielectric properties of blood cells. Lateral magnetophoretic (MAP) force is achieved using a high-gradient magnetic field, caused by a ferromagnetic wire array inlaid on the bottom glass substrate. Lateral dielectrophoretic (DEP) force is achieved using a planar interdigitated electrode array, patterned on the top glass substrate. Red blood cells in peripheral blood are primarily driven by the lateral MAP force, while white blood cells are primarily forced by the lateral DEP force, the direction of which is opposite to that of the lateral MAP force. These lateral MAP and DEP forces are produced evenly on the whole area of the microchannel, thereby achieving highly efficient enrichment. The experimental results showed that the lateral MAP-DEP microseparator can continuously enrich circulating nucleated cells by 20,000-fold from peripheral blood simply by using an external magnetic flux of 0.3 T and a 2-MHz sinusoidal voltage of 4 Vp-p. Additionally, by using the intrinsic magnetic and dielectric properties of blood cells, we eliminated the need for laborious sample preparation procedures before and after enrichment, and also reduced the cost.     

Analytical model of microfluidic transport of non-magnetic particles in ferrofluids under the influence of a permanent magnet

This study describes an analytical model and experimental verifications of transport of non-magnetic spherical microparticles in ferrofluids in a microfluidic system that consists of a microchannel and a permanent magnet. The permanent magnet produces a spatially non-uniform magnetic field that gives rise to a magnetic buoyancy force on particles within ferrofluid-filled microchannel. We obtained trajectories of particles in the microchannel by (1) calculating magnetic buoyancy force through the use of an analytical expression of magnetic field distributions and a nonlinear magnetization model of ferrofluids, (2) deriving governing equations of motion for particles through the use of analytical expressions of dominant magnetic buoyancy and hydrodynamic viscous drag forces, (3) solving equations of motion for particles in laminar flow conditions. We studied effects of particle size and flow rate in the microchannel on the trajectories of particles. The analysis indicated that particles were increasingly deflected in the direction that was perpendicular to the flow when size of particles increased, or when flow rate in the microchannel decreased. We also studied ??wall effect?? on the trajectories of particles in the microchannel when surfaces of particles were in contact with channel wall. Experimentally obtained trajectories of particles were used to confirm the validity of our analytical results. We believe this study forms the theoretical foundation for size-based particle (both synthetic and biological) separation in ferrofluids in a microfluidic device. The simplicity and versatility of our analytical model make it useful for quick optimizations of future separation devices as the model takes into account important design parameters including particle size, property of ferrofluids, magnetic field distribution, dimension of microchannel, and fluid flow rate.     

Improved particle concentration by cascade AC electroosmotic flow

The importance of electrokinetics in microfluidic technology has been growing owing to its versatility and simplicity in fabrication, implementation, and handling. Alternating-current electroosmosis (ACEO), which is the motion of fluid due to the ion movement by an interaction between AC electric field and an electrical double layer on the electrode surface, has a potential for a particle concentration method to detecti rare samples flowing in a microchannel. This study investigates an improved ACEO-based particle concentration by cascade electrokinetic approach. Flow field induced by ACEO and accumulation behavior of particles were parametrically measured to discuss the concentrating mechanism. The accumulation of particles by ACEO can be explained by a balance between the attenuating electroosmotic flow to transport particles and the inherent diffusive motion of the particles, which is hindered due to the near-wall location. Although a parallel double-gap electrode geometry enables particles to be collected at the center of electrode very sharply, it has scattering zones with accumulated particles at sidewalls of the channel. This drawback can be overcome by applying sheath flow or introducing cascade electrode pattern upstream of the focusing zone. As a result, total concentration efficiency was 98.4 % for all the particles flowing in the cascade device. The resultant concentrated particles exist on the electrode surface within 5 μm, and three-dimensional concentration of particle with the concentration factor as large as 700 is possible using a monolithic channel, co-planar electrode, and sheathless solution feeding. This cascade electrokinetic method provides a new and effective preconcentrator for ultra-sensitive detection of rare samples.     

On-chip whole blood plasma separator based on microfiltration,sedimentation and wetting contrast

Miniaturized on-chip blood separators have a great value for point-of-care diagnosis. In our work, a combined design strategy—microfiltration, sedimentation in a retarded flow, and wetting contrast—was taken to overcome the known limitations of on-chip blood separators. Our microfluidic chip consists of a polydimethylsiloxane micropillar array and an etched glass with microchannel branches. The red blood cells are significantly slowed and gradually settled down due to micropillars and enlarged dimension of a chamber. An etched glass microchannel allows the extraction of blood plasma exclusively due to the capillary effect. The fabricated microfluidic device can separate blood plasma from a whole blood sample without any external driving force or dilution. The measured plasma separation efficiency was close to 100 % from human whole blood. Autonomous on-chip separation and collection of blood plasma was demonstrated.     

Multiplex Inertio-Magnetic Fractionation (MIMF) of magnetic and non-magnetic microparticles in a microfluidic device

Separation of multiple microparticles at high throughput is highly required in different applications such as diagnostics and immunomagnetic detection. We present a microfluidic device for multiplex (i.e., duplex to fourplex) fractionation of magnetic and non-magnetic microparticles using a novel hybrid technique based on interactions between flow-induced inertial forces and countering magnetic forces in a simple expansion microchannel with a side permanent magnet. Separation of more than two types of particles solely by inertia or magnetic forces in a straight microchannel is challenging due to the inherent limitations of each technique. By combining inertial and magnetic forces in a straight microchannel and addition of a downstream expansion hydrodynamic separator, we overcame these limitations and achieved duplex to fourplex fractionation of magnetic and non-magnetic microparticles with high throughput and efficiency. Particle fractionation performance in our device was first optimized with respect to parameters such as flow rate and aspect ratio of the channel to attain coexistence of inertial and magnetic focusing of particles. Using this scheme, we achieved duplex fractionation of particles at high throughput of 10⁹ particles per hour. Further, we conducted experiments with three magnetic particles (5, 11 and 35 µm) to establish their size-dependent ordering in the device under combined effects of magnetic and inertial forces. We then used the findings for fourplex fractionation of 5, 11 and 35 µm magnetic particles from non-magnetic particles of various sizes (10–19 µm). This Multiplex Inertio-Magnetic Fractionation (MIMF) technique offers a simple tool to handle complex and heterogeneous samples and can be used for affinity-based immunomagnetic separation of multiple biological substances in fluidic specimens in the future.     

Experimental study on self-flowing speed in microchannel related to micro-/nanoscale surface topographies

The microfluidic flowing on chip surface depends on the external load such as centrifugal force, magnetic force and bubbles, but it leads to the complexity of microsystem. Hence, a self-flowing is proposed inside microchannel on chip surface without any external load. The objective is to explore how micro-/nanoscale surface topographies of microchannel influence the microfluidic flowing. First, the microgrinding with a diamond wheel microtip was employed to fabricate the accurate and smooth V-shaped microchannel with the height of 300 µm and less; then, the microfluidic flowing state was modeled by the flowing concave with the parameterization of flow-end height; finally, the microfluidic flowing speed was experimentally investigated with reference to microchannel angle, gradient, surface roughness and chip material. It is shown that the microfluidic self-flowing is mainly induced by the microchannel tip and the nanometer-scale surface cracks around the microchannel tip. Small microchannel angle, large microchannel gradient and smooth microchannel surface may enhance the flowing speed on chip surface. The brittle quartz glass produces the nanometer-scale surface cracks around the microchannel tip, leading to an increase about 40 times in the self-flowing speed compared with the ductile polymer. It is confirmed that the self-flowing speed in dynamic state may be characterized by the proposed concave flow-end height.