High-throughput inertial particle focusing in a curved microchannel: Insights into the flow-rate regulation mechanism and process model

In this work, we design and fabricate a miniaturized spiral-shaped microchannel device which can be used for high-throughput particle/cell ordering, enrichment, and purification. To probe into the flow rate regulation mechanism, an experimental investigation is carried out on the focusing behaviors of particles with significantly different sizes in this device. A complete picture of the focusing position shifting process is unfolded to clarify the confusing results obtained from flow regimes with different dominant forces in past research. Specifically, with the increase of the flow rate, particles are observed to first move towards the inner wall under the dominant inertial migration, then stabilize at a specific position and finally shift away from the inner wall due to the alternation of the dominant force. Novel phenomena of focusing instability, co-focusing, and focusing position interchange of differently sized particles are also observed and investigated. Based on the obtained experimental data, we develop and validate, for the first time, a five-stage model of the particle focusing process with increasing flow rate for interpreting particle behaviors in terms of the competition between inertial lift and Dean drag forces. These new experimental findings and the proposed process model provide an important supplement to the existing mechanism of inertial particle flow and enable more flexible and precise particle manipulation. Additionally, we examine the focusing behaviors of bioparticles with a polydisperse size distribution to validate the explored mechanisms and thus help realize efficient enrichment and purification of these particles.     

Inertial microfluidics in contraction–expansion microchannels: A review

Inertial microfluidics has brought enormous changes in the conventional cell/particle detection process and now become the main trend of sample pretreatment with outstanding throughput, low cost, and simple control method. However, inertial microfluidics in a straight microchannel is not enough to provide high efficiency and satisfying performance for cell/particle separation. A contraction–expansion microchannel is a widely used and multifunctional channel pattern involving inertial microfluidics, secondary flow, and the vortex in the chamber. The strengthened inertial microfluidics can help us to focus particles with a shorter channel length and less processing time. Both the vortex in the chamber and the secondary flow in the main channel can trap the target particles or separate particles based on their sizes more precisely. The contraction–expansion microchannels are also capable of combining with a curved, spiral, or serpentine channel to further improve the separation performance. Some recent studies have focused on the viscoelastic fluid that utilizes both elastic forces and inertial forces to separate different size particles precisely with a relatively low flow rate for the vulnerable cells. This article comprehensively reviews various contraction–expansion microchannels with Newtonian and viscoelastic fluids for particle focusing, separation, and microfluid mixing and provides particle manipulation performance data analysis for the contraction–expansion microchannel design.     

Three-dimensional cellular focusing utilizing a combination of insulator-based and metallic dielectrophoresis

Particle focusing in microfluidic devices is a necessary step in medical applications, such as detection, sorting, counting, and flow cytometry. This study proposes a microdevice that combines insulator-based and metal-electrode dielectrophoresis for the three-dimensional focusing of biological cells. Four insulating structures, which form an X pattern, are employed to confine the electric field in a conducting solution, thereby creating localized field minima in the microchannel. These electrodes, 56-μm-wide at the top and bottom surfaces, are connected to one electric pole of the power source. The electrodes connected to the opposite pole, which are at the sides of the microchannel, have one of three patterns: planar, dual-planar, or three-dimensional. Therefore, low-electric-field regions at the center of the microchannel are generated to restrain the viable HeLa cells with negative dielectrophoretic response. The array of insulating structures aforementioned is used to enhance the performance of confinement. According to numerical simulations, three-dimensional electrodes exhibit the best focusing performance, followed by dual-planar and planar electrodes. Experimental results reveal that increasing the strength of the applied electric field or decreasing the inlet flow rate significantly enhances focusing performance. The smallest width of focusing is 17 μm for an applied voltage and an inlet flow rate of 35 V and 0.5 μl/min, respectively. The effect of the inlet flow rate on focusing is insignificant for an applied voltage of 35 V. The proposed design retains the advantages of insulator-based dielectrophoresis with a relatively low required voltage. Additionally, complicated flow controls are unnecessary for the three-dimensional focusing of cells.     

Dean-flow-coupled elasto-inertial three-dimensional particle focusing under viscoelastic flow in a straight channel with asymmetrical expansion–contraction cavity arrays

In this paper, 3D particle focusing in a straight channel with asymmetrical expansion–contraction cavity arrays (ECCA channel) is achieved by exploiting the dean-flow-coupled elasto-inertial effects. First, the mechanism of particle focusing in both Newtonian and non-Newtonian fluids was introduced. Then particle focusing was demonstrated experimentally in this channel with Newtonian and non-Newtonian fluids using three different sized particles (3.2 μm, 4.8 μm, and 13 μm), respectively. Also, the effects of dean flow (or secondary flow) induced by expansion–contraction cavity arrays were highlighted by comparing the particle distributions in a single straight rectangular channel with that in the ECCA channel. Finally, the influences of flow rates and distances from the inlet on focusing performance in the ECCA channel were studied. The results show that in the ECCA channel particles are focused on the cavity side in Newtonian fluid due to the synthesis effects of inertial and dean-drag force, whereas the particles are focused on the opposite cavity side in non-Newtonian fluid due to the addition of viscoelastic force. Compared with the focusing performance in Newtonian fluid, the particles are more easily and better focused in non-Newtonian fluid. Besides, the Dean flow in visco-elastic fluid in the ECCA channel improves the particle focusing performance compared with that in a straight channel. A further advantage is three-dimensional (3D) particle focusing that in non-Newtonian fluid is realized according to the lateral side view of the channel while only two-dimensional (2D) particle focusing can be achieved in Newtonian fluid. Conclusively, this novel Dean-flow-coupled elasto-inertial microfluidic device could offer a continuous, sheathless, and high throughput (>10 000 s⁻¹) 3D focusing performance, which may be valuable in various applications from high speed flow cytometry to cell counting, sorting, and analysis.     

High-performance microfluidic rectifier based on sudden expansion channel with embedded block structure

A high-performance microfluidic rectifier incorporating a microchannel and a sudden expansion channel is proposed. In the proposed device, a block structure embedded within the expansion channel is used to induce two vortex structures at the end of the microchannel under reverse flow conditions. The vortices reduce the hydraulic diameter of the microchannel and, therefore, increase the flow resistance. The rectification performance of the proposed device is evaluated by both experimentally and numerically. The experimental and numerical values of the rectification performance index (i.e., the diodicity, Di) are found to be 1.54 and 1.76, respectively. Significantly, flow rectification is achieved without the need for moving parts. Thus, the proposed device is ideally suited to the high pressure environment characteristic of most micro-electro-mechanical-systems (MEMS)-based devices. Moreover, the rectification performance of the proposed device is superior to that of existing valveless rectifiers based on Tesla valves, simple nozzle/diffuser structures, or cascaded nozzle/diffuser structures.     

Continuous size-based separation of microparticles in a microchannel with symmetric sharp corner structures

A new microchannel with a series of symmetric sharp corner structures is reported for passive size-dependent particle separation. Micro particles of different sizes can be completely separated based on the combination of the inertial lift force and the centrifugal force induced by the sharp corner structures in the microchannel. At appropriate flow rate and Reynolds number, the centrifugal force effect on large particles, induced by the sharp corner structures, is stronger than that on small particles; hence after passing a series of symmetric sharp corner structures, large particles are focused to the center of the microchannel, while small particles are focused at two particle streams near the two side walls of the microchannel. Particles of different sizes can then be completely separated. Particle separation with this device was demonstrated using 7.32 μm and 15.5 μm micro particles. Experiments show that in comparison with the prior multi-orifice flow fractionation microchannel and multistage-multiorifice flow fractionation microchannel, this device can completely separate two-size particles with narrower particle stream band and larger separation distance between particle streams. In addition, it requires no sheath flow and complex multi-stage separation structures, avoiding the dilution of analyte sample and complex operations. The device has potentials to be used for continuous, complete particle separation in a variety of lab-on-a-chip and biomedical applications.     

An insulator-based dielectrophoretic microdevice for the simultaneous filtration and focusing of biological cells

Manipulating and discriminating biological cells of interest using microfluidic and micro total analysis system (μTAS) devices have potential applications in clinical diagnosis and medicine. Cellular focusing in microfluidic devices is a prerequisite for medical applications, such as cell sorting, cell counting, or flow cytometry. In the present study, an insulator-based dielectrophoretic microdevice is designed for the simultaneous filtration and focusing of biological cells. The cells are introduced into the microchannel and hydrodynamically pre-confined by funnel-shaped insulating structures close to the inlet. There are ten sets of X-patterned insulating structures in the microfluidic channel. The main function of the first five sets of insulating structures is to guide the cells by negative dielectrophoretic responses (viable HeLa cells) into the center region of the microchannel. The positive dielectrophoretic cells (dead HeLa cells) are attracted to regions with a high electric-field gradient generated at the edges of the insulating structures. The remaining five sets of insulating structures are mainly used to focus negative dielectrophoretic cells that have escaped from the upstream region. Experiments employing a mixture of dead and viable HeLa cells are conducted to demonstrate the effectiveness of the proposed design. The results indicate that the performance of both filtration and focusing improves with the increasing strength of the applied electric field and a decreasing inlet sample flow rate, which agrees with the trend predicted by the numerical simulations. The filtration efficiency, which is quantitatively investigated, is up to 88% at an applied voltage of 50 V peak-to-peak (1 kHz) and a sample flow rate of 0.5 μl/min. The proposed device can focus viable cells into a single file using a voltage of 35 V peak-to-peak (1 kHz) at a sample flow rate of 1.0 μl/min.     

High-throughput particle separation and concentration using spiral inertial filtration

A spiral inertial filtration (SIFT) device that is capable of high-throughput (1 ml/min), high-purity particle separation while concentrating recovered target particles by more than an order of magnitude is reported. This device is able to remove large fractions of sample fluid from a microchannel without disruption of concentrated particle streams by taking advantage of particle focusing in inertial spiral microfluidics, which is achieved by balancing inertial lift forces and Dean drag forces. To enable the calculation of channel geometries in the SIFT microsystem for specific concentration factors, an equivalent circuit model was developed and experimentally validated. Large particle concentration factors were then achieved by maintaining either the average fluid velocity or the Dean number throughout the entire length of the channel during the incremental removal of sample fluid. The SIFT device was able to separate MCF7 cells spiked into whole blood from the non-target white blood cells (WBC) with a recovery of nearly 100% while removing 93% of the sample volume, which resulted in a concentration enhancement of the MCF7 cancer cells by a factor of 14.     

Thermal mixing of two miscible fluids in a T-shaped microchannel

In this paper, thermal mixing characteristics of two miscible fluids in a T-shaped microchannel are investigated theoretically, experimentally, and numerically. Thermal mixing processes in a T-shaped microchannel are divided into two zones, consisting of a T-junction and a mixing channel. An analytical two-dimensional model was first built to describe the heat transfer processes in the mixing channel. In the experiments, de-ionized water was employed as the working fluid. Laser induced fluorescence method was used to measure the fluid temperature field in the microchannel. Different combinations of flow rate ratios were studied to investigate the thermal mixing characteristics in the microchannel. At the T-junction, thermal diffusion is found to be dominant in this area due to the striation in the temperature contours. In the mixing channel, heat transfer processes are found to be controlled by thermal diffusion and convection. Measured temperature profiles at the T-junction and mixing channel are compared with analytical model and numerical simulation, respectively.     

Microfluidic on-chip fluorescence-activated interface control system

A microfluidic dynamic fluorescence-activated interface control system was developed for lab-on-a-chip applications. The system consists of a straight rectangular microchannel, a fluorescence excitation source, a detection sensor, a signal conversion circuit, and a high-voltage feedback system. Aqueous NaCl as conducting fluid and aqueous glycerol as nonconducting fluid were introduced to flow side by side into the straight rectangular microchannel. Fluorescent dye was added to the aqueous NaCl to work as a signal representing the interface position. Automatic control of the liquid interface was achieved by controlling the electroosmotic effect that exists only in the conducting fluid using a high-voltage feedback system. A LABVIEW program was developed to control the output of high-voltage power supply according the actual interface position, and then the interface position is modified as the output of high-voltage power supply. At last, the interface can be moved to the desired position automatically using this feedback system. The results show that the system presented in this paper can control an arbitrary interface location in real time. The effects of viscosity ratio, flow rates, and polarity of electric field were discussed. This technique can be extended to switch the sample flow and droplets automatically.     

An efficient micromixer based on multidirectional vortices due to baffles and channel curvature

An efficient planar micromixer based on multidirectional vortices in a curved channel with radial baffles is proposed and examined in this work. The curvature of the microchannel and the radial baffles induce vortices in different directions. The multidirectional vortices and the converging-diverging flow caused by the baffles contribute together to the enhancement of mixing. The micromixer is fabricated with polydimethylsiloxane by a single planar microlithography process and the mixing behaviors are observed by a confocal spectral microscope imaging system to validate the simulation obtained by a commercial code. The simulation and experimental results are in reasonable agreement. The concentration distributions and flow patterns obtained reveal the following trends. (i) The mixing efficiency of the basic C-shaped micromixer with the first baffle attached to the internal cylinder and the second attached to the external cylinder is better than that of the C-shaped micromixer with inverted arrangement of baffles. (ii) When the radius of the curved channel and the width of the passage between the baffle and the cylindrical wall are small enough and the Reynolds number (Re) is large enough, an extra separation vortex develops in the downstream of the second baffle. This phenomenon is one of the reasons of trend (i). (iii) A micromixer consisting of a few basic C-shaped micromixers connected by straight channels may generate a high degree of mixing for the case with a large Re.     

Effect of electrical double layer on electric conductivity and pressure drop in a pressure-driven microchannel flow

The effect of an electrical double layer (EDL) on microchannel flow has been studied widely, and a constant bulk electric conductivity is often used in calculations of flow rate or pressure drop. In our experimental study of pressure-driven micropipette flows, the pipette diameter is on the same order of magnitude as the Debye length. The overlapping EDL resulted in a much higher electric conductivity, lower streaming potential, and lower electroviscous effect. To elucidate the effect of overlapping EDL, this paper developed a simple model for water flow without salts or dissolved gases (such as CO₂) inside a two-dimensional microchannel. The governing equations for the flow, the Poisson, and Nernst equations for the electric potential and ion concentrations and the charge continuity equation were solved. The effects of overlapping EDL on the electric conductivity, velocity distribution, and overall pressure drop in the microchannel were quantified. The results showed that the average electric conductivity of electrolyte inside the channel increased significantly as the EDL overlaps. With the modified mean electric conductivity, the pressure drop for the pressure-driven flow was smaller than that without the influence of the EDL on conductivity. The results of this study provide a physical explanation for the observed decrease in electroviscous effect for microchannels when the EDL layers from opposing walls overlap.     

并联管组模型流动压损分析（一）

流体在并联管组模型中流动的均匀性问题具有非常广泛的应用背景。U型分布的是循环电液电堆中经常使用的模型。通过Fluent软件对其进行数值模拟,试图探讨单体个数和电堆压损之间的关系。经计算发现,在流量一定的前提下,随单体数量的增加,电堆压损递减,但不是单调递减,而是存在一个关键的拐点,当单体数超过拐点值后,压损基本恒定。另通过模拟计算发现,在单体数一定的情况下,电堆压损与流量对应关系。     

Microfluidic based high throughput synthesis of lipid-polymer hybrid nanoparticles with tunable diameters

Core-shell hybrid nanoparticles (NPs) for drug delivery have attracted numerous attentions due to their enhanced therapeutic efficacy and good biocompatibility. In this work, we fabricate a two-stage microfluidic chip to implement a high-throughput, one-step, and size-tunable synthesis of mono-disperse lipid-poly (lactic-co-glycolic acid) NPs. The size of hybrid NPs is tunable by varying the flow rates inside the two-stage microfluidic chip. To elucidate the mechanism of size-controllable generation of hybrid NPs, we observe the flow field in the microchannel with confocal microscope and perform the simulation by a numerical model. Both the experimental and numerical results indicate an enhanced mixing effect at high flow rate, thus resulting in the assembly of small and mono-disperse hybrid NPs. In vitro experiments show that the large hybrid NPs are more likely to be aggregated in serum and exhibit a lower cellular uptake efficacy than the small ones. This microfluidic chip shows great promise as a robust platform for optimization of nano drug delivery system.     

Deformation properties between fluid and periodic circular obstacles in polydimethylsiloxane microchannels: Experimental and numerical investigations under various conditions

Understanding the mechanical properties of optically transparent polydimethylsiloxane (PDMS) microchannels was essential to the design of polymer-based microdevices. In this experiment, PDMS microchannels were filled with a 100 μM solution of rhodamine 6G dye at very low Reynolds numbers (∼10⁻³). The deformation of PDMS microchannels created by pressure-driven flow was investigated by fluorescence microscopy and quantified the deformation by the linear relationship between dye layer thickness and intensity. A line scan across the channel determined the microchannel deformation at several channel positions. Scaling analysis widely used to justify PDMS bulging approximation was allowed when the applied flow rate was as high as 2.0 μl/min. The three physical parameters (i.e., flow rate, PDMS wall thickness, and mixing ratio) and the design parameter (i.e., channel aspect ratio = channel height/channel width) were considered as critical parameters and provided the different features of pressure distributions within polymer-based microchannel devices. The investigations of the four parameters performed on flexible materials were carried out by comparison of experiment and finite element method (FEM) results. The measured Young''s modulus from PDMS tensile test specimens at various circumstances provided reliable results for the finite element method. A thin channel wall, less cross-linker, high flow rate, and low aspect ratio microchannel were inclined to have a significant PDMS bulging. Among them, various mixing ratios related to material property and aspect ratios were one of the significant factors to determine PDMS bulging properties. The measured deformations were larger than the numerical simulation but were within corresponding values predicted by the finite element method in most cases.     

Hydrodynamic self-focusing in a parallel microfluidic device through cross-filtration

The flow focusing is a fundamental prior step in order to sort, analyze, and detect particles or cells. The standard hydrodynamic approach requires two fluids to be injected into the microfluidic device: one containing the sample and the other one, called the sheath fluid, allows squeezing the sample fluid into a narrow stream. The major drawback of this approach is the high complexity of the layout for microfluidic devices when parallel streams are required. In this work, we present a novel parallelized microfluidic device that enables hydrodynamic focusing in each microchannel using a single feed flow. At each of the parallel channels, a cross-filter region is present that allows removing fluid from the sample fluid. This fluid is used to create local sheath fluids that hydrodynamically pinch the sample fluid. The great advantage of the proposed device is that, since only one inlet is needed, multiple parallel micro-channels can be easily introduced into the design. In the paper, the design method is described and the numerical simulations performed to define the optimal design are summarized. Moreover, the operational functionality of devices tested by using both polystyrene beads and Acute Lymphoid Leukemia cells are shown.     

Migration distance-based platelet function analysis in a microfluidic system

Aggregation and adhesion of platelets to the vascular wall are shear-dependent processes that play critical roles in hemostasis and thrombosis at vascular injury sites. In this study, we designed a simple and rapid assay of platelet aggregation and adhesion in a microfluidic system. A shearing mechanism using a rotating stirrer provided adjustable shear rate and shearing time and induced platelet activation. When sheared blood was driven through the microchannel under vacuum pressure, shear-activated platelets adhered to a collagen-coated surface, causing blood flow to significantly slow and eventually stop. To measure platelet adhesion and aggregation, the migration distance (MD) of blood through the microchannel was monitored. As the microstirrer speed increased, MD initially decreased exponentially but then increased beyond a critical rpm. For platelet-excluded blood samples, there were no changes in MD with increasing stirrer speed. These findings imply that the stirrer provided sufficiently high shear to activate platelets and that blood MD is a potentially valuable index for measuring the shear-dependence of platelet activation. Our microfluidic system is quick and simple, while providing a precise assay to measure the effects of shear on platelet aggregation and adhesion.     

Mixing enhancement in T-junction microchannel with acoustic streaming induced by triangular structure

The T-shaped microchannel system is used to mix similar or different fluids, and the laminar flow nature makes the mixing at the entrance junction region a challenging task. Acoustic streaming is a steady vortical flow phenomenon that can be produced in the microchannel by oscillating acoustic transducer around the sharp edge tip structure. In this study, the acoustic streaming is produced using a triangular structure with tip angles of 22.62°, 33.4°, and 61.91°, which is placed at the entrance junction region and mixes the inlets flow from two directions. The acoustic streaming flow patterns were investigated using micro-particle image velocimetry (μPIV) in various tip edge angles, flow rate, oscillation frequency, and amplitude. The velocity and vorticity profiles show that a pair of counter-rotating streaming vortices were created around the sharp triangle structure and raised the Z vorticity up to 10 times more than the case without acoustic streaming. The mixing experiments were performed by using fluorescent green dye solution and de-ionized water and evaluated its performance with the degree of mixing (M) at different amplitudes, flow rates, frequencies, and tip edge angles using the grayscale value of pixel intensity. The degree of mixing characterized was found significantly improved to 0.769 with acoustic streaming from 0.4017 without acoustic streaming, in the case of 0.008 μl/min flow rate and 38 V oscillation amplitude at y = 2.15 mm. The results suggested that the creation of acoustic streaming around the entrance junction region promotes the mixing of two fluids inside the microchannel, which is restricted by the laminar flow conditions.     

A numerical study on distributions during cryoprotectant loading caused by laminar flow in a microchannel

In this work, we conduct a computational study on the loading of cryoprotective agents into cells in preparation for cryopreservation. The advantages of microfluidics in cryopreserving cells include control of fluid flow parameters for reliable cryoprotectant loading and reproducible streamlined processing of samples. A 0.25 m long, three inlet T-junction microchannel serves as an idealized environment for this process. The flow field and concentration distribution are determined from a computational fluid dynamics study and cells are tracked as inert particles in a Lagrangian frame. These particles are not confined to streamlines but can migrate laterally due to the Segre-Sildeberg effect for particles in a shear flow. During this tracking, the local concentration field surrounding the cell is monitored. This data are used as input into the Kedem-Katchalsky equations to numerically study passive solute transport across the cell membrane. As a result of the laminar flow, each cell has a unique pathline in the flow field resulting in different residence times and a unique external concentration field along its path. However, in most previous studies, the effect of a spatially varying concentration field on the transport across the cell membrane is ignored. The dynamics of this process are investigated for a population of cells released from the inlet. Using dimensional analysis, we find a governing parameter α, which is the ratio of the time scale for membrane transport to the average residence time in the channel. For α <  = 0.224, cryoprotectant loading is completed to within 5% of the target concentration for all of the cells. However, for α > 0.224, we find the population of cells does not achieve complete loading and there is a distribution of intracellular cryoprotective agent concentration amongst the population. Further increasing α beyond a value of 2 leads to negligible cryoprotectant loading. These simulations on populations of cells may lead to improved microfluidic cryopreservation protocols where more consistent cryoprotective agent loading and freezing can be achieved, thus increasing cell survival.