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.     

Optimizing micromixer design for enhancing dielectrophoretic microconcentrator performance

We present an investigation into optimizing micromixer design for enhancing dielectrophoretic (DEP) microconcentrator performance. DEP-based microconcentrators use the dielectrophoretic force to collect particles on electrodes. Because the DEP force generated by electrodes decays rapidly away from the electrodes, DEP-based microconcentrators are only effective at capturing particles from a limited cross section of the input liquid stream. Adding a mixer can circulate the input liquid, increasing the probability that particles will drift near the electrodes for capture. Because mixers for DEP-based microconcentrators aim to circulate particles, rather than mix two species, design specifications for such mixers may be significantly different from that for conventional mixers. Here we investigated the performance of patterned-groove micromixers on particle trapping efficiency in DEP-based microconcentrators numerically and experimentally. We used modeling software to simulate the particle motion due to various forces on the particle (DEP, hydrodynamic, etc.), allowing us to predict trapping efficiency. We also conducted trapping experiments and measured the capture efficiency of different micromixer configurations, including the slanted groove, staggered herringbone, and herringbone mixers. Finally, we used these analyses to illustrate the design principles of mixers for DEP-based concentrators.     

Continuous sorting of microparticles using dielectrophoresis

Sorting of particles such as cells is a critical process for many biomedical applications, and it is challenging to integrate it into an analytical microdevice. We report an effective and flexible dielectrophoresis (DEP)-based microfluidic device for continuous sorting of multiple particles in a microchannel. The particle sorter is composed of two components-a DEP focusing unit and a Movable DEP Trap (MDT). The trap is formed by an array of microelectrodes at the bottom of the channel and a transparent electrode plate placed at the top. The location of the trap is dependent on the configuration of voltages on the array and therefore is addressable. Flowing particles are first directed and focused into a single particle stream by the focusing unit. The streamed particles are then sorted into different fractions using the movable trap by rapidly switching the applied voltage. The performance of the sorter is demonstrated by successfully sorting microparticles in a continuous flow. The proposed DEP-based microfluidic sorter can be implemented in applications such as sample preparation and cell sorting for subsequent analytical processing, where sorting of particles is needed.     

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.     

Dielectrophoresis in microchips containing arrays of insulating posts: theoretical and experimental results

Dielectrophoresis (DEP), a nonlinear electrokinetic transport mechanism, can be used to concentrate and sort cells, viruses, and particles. To date, microfabricated DEP-based devices have typically used embedded metal electrodes to apply spatially nonuniform, time-varying (AC) electric fields. We have developed an alternative method in which arrays of insulating posts in a channel of a microchip produce the spatially nonuniform fields needed for DEP. Electrodes may be located remotely, allowing operation of the device down to zero frequency (DC) without excessive problems of electrolysis. Applying a sufficiently large electric field across an insulating-post array produces two flow regimes through a competition between electrokinetic flow (combined electrophoresis and electroosmosis) and dielectrophoresis. "Streaming DEP" is observed when DEP dominates diffusion but is overcome by electrokinetic flow. Particles concentrated by DEP forces in areas of electric field extrema travel electrokinetically down the array in flowing streams. In an array of posts, dielectrophoretic forcing within repeated rows adds coherently to produce flowing streams of highly concentrated and rarefied particles. We demonstrate that this reinforcement is a strong function of alignment of the array with respect to the applied electric field and that the particle concentrations can be "enhanced" or "depleted" along columns of posts, enabling a novel class of continuous-flow, selective particle filter/concentrator devices. To our knowledge, this is the first observation of streaming dielectrophoresis. The second regime is "trapping DEP," in which DEP forces dominate over both diffusion and electrokinetic flow, reversibly immobilizing particles on the insulating posts, enabling inexpensive and embedded batch filter/concentrator devices. Devices can be biased electrically to manipulate particles selectively by varying the field strength to vary the relative magnitudes of electrokinetic flow and DEP. Post shapes are contoured easily to control electric field gradients and, hence, DEP behavior. Simple simulations based on similitude of fluid flow and electric field that solve the Laplace equation to obtain fluid velocity have also been developed to predict the dielectrophoretic behavior in an array of posts. These simulations are in excellent agreement with the experimental observations and provide insight into electrokinetic behavior to enable design of dielectrophoretic concentrators and sorters.     

Effect of particle-particle interaction on dielectrophoretic single particle trap in a sudden contraction flow

Dielectrophoretic(DEP) force is significant in manipulating tiny objects in micro/nano scale. To study the effect of electric interaction force on particle manipulation, a microstructure consisting of a pair of strip electrodes and a sudden contraction micro-channel was constructed. Besides DEP force and hydrodynamic force acting on single particle, the numerical model also involved electric interaction force and force moment on two particles. The analyses revealed that the particle-particle interaction force was in the same order as that of DEP force on single trapped particle. The interaction force resulted in trapping single particle failure under continuous DEP force.Thus, pulsed DEP force, turning on/off DEP force at a given time interval, was suggested. During the "off" period,the velocity difference of the two particles located at sudden contraction micro-channel enlarged the gap between them and further weakened the particle-particle interaction. By a proof-of-concept experiment, both the trapping behavior of single particle and that of two particles were in good agreement with the model.With carefully controlled parameters, the reliable function of retaining single particle was realized by pulsed DEP.     

On-chip high-speed sorting of micron-sized particles for high-throughput analysis

A new design of particle sorting chip is presented. The device employs a dielectrophoretic gate that deflects particles into one of two microfluidic channels at high speed. The device operates by focussing particles into the central streamline of the main flow channel using dielectrophoretic focussing. At the sorting junction (T- or Y-junction) two sets of electrodes produce a small dielectrophoretic force that pushes the particle into one or other of the outlet channels, where they are carried under the pressure-driven fluid flow to the outlet. For a 40 microm wide and high channel, it is shown that 6 microm diameter particles can be deflected at a rate of 300/s. The principle of a fully automated sorting device is demonstrated by separating fluorescent from non-fluorescent latex beads.     

High-throughput and high-resolution flow cytometry in molded microfluidic devices

We describe the design, fabrication, and operation of two types of flow cytometers based on microfluidic devices made of a single cast of poly(dimethylsiloxane). The stream of particles or cells injected into the devices is hydrodynamically focused in both transverse and lateral directions, has a uniform velocity, and has adjustable diameter and shape. The cytometry system built around the first microfluidic device has fluorescence detection accuracy comparable with that of a commercial flow cytometer and can analyze as many as 17 000 particles/s. This high-throughput microfluidic device could be used in inexpensive stand-alone cytometers or as a part of integrated microanalysis systems. In the second device, a stream of particles is focused to a flow layer of a submicrometer thickness that allows imaging the particles with a high numerical aperture microscope objective. To take long-exposure, low-light fluorescence images of live cells, the device is placed on a moving stage, which accurately balances the translational motion of particles in the flow. The achieved resolution is comparable to that of still micrographs. This high-resolution device could be used for analysis of morphology and fluorescence distribution in cells in continuous flow.     

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.     

Continuous-flow particle separation by 3D Insulative dielectrophoresis using coherently shaped, dc-biased, ac electric fields

We present the development of a continuous-flow, "dielectrophoretic spectrometer" based on insulative DEP techniques and three-dimensional geometric design. Hot-embossed thermoplastic devices allow for high-throughput analysis and geometric control of electric fields via ridged microstructures patterned in a high width-to-depth aspect ratio (250:1) channel. We manipulate particles with dc-biased, ac electric fields and generate continuous-output streams of particles with a transverse outlet position specified by linear and nonlinear particle mobilities. We show, with simulation and experiment, that characteristic shape factors can be defined that capture the effects of constrictions in channel depth and that modulating the angle of these constrictions changes the resulting local DEP force. Microdevices are fabricated with an insulative constriction in channel depth, whose angle of incidence with the direction of flow varies continuously across the channel width. The resulting electric field gradients enable demonstration of a dielectrophoretic spectrometer that separates particles and controls their transverse channel position.     

Sheathless hydrophoretic particle focusing in a microchannel with exponentially increasing obstacle arrays

We present a novel microfluidic device with exponentially increasing obstacle arrays to enable sheathless particle focusing. The anisotropic fluidic resistance of slant obstacles generates transverse flows, along which particles are focused to one sidewall. In the successive channel with exponentially increasing widths, bent obstacles extended from the slant obstacles increase the focusing efficiency of the particles. With the device, we achieved the focusing efficiency of 76%, 94%, and 98% for 6, 10, and 15 microm beads, respectively. The focusing efficiency of the particles can be further improved in the devices with more extension steps. In addition, using the microfluidic devices with the symmetric structure of the slant and bent obstacles, we achieved complete focusing of biological cells to the centerline of a channel within 1.7% coefficient of variation. The results demonstrated the sheathless hydrophoretic focusing of microparticles and cells with the advantages of a sheathless method, passive operation, single channel, and flow rate independence.     

Dielectrophoresis-activated multiwell plate for label-free high-throughput drug assessment

Dielectrophoresis (DEP) offers many advantages over conventional cell assays such as flow cytometry and patch clamp techniques for assessing cell electrophysiology as a marker for cancer studies and drug interaction assessment. However, despite the advantages offered by DEP analysis, uptake has been low, remaining largely in the academic arena, due to the process of analysis being time-consuming, laborious, and ultimately allowing only serial analysis on small numbers of cells. In this paper we describe a new method of performing DEP analysis based on laminate manufacturing methods. These use a three-dimensional "well" structure, similar in size and pitch to conventional microtiter well plates, but offer electrodes along the inner surface to allow easy measurement of cell properties through the whole population. The result can then be determined rapidly using a conventional well-plate reader. The nature of the device means that many electrodes, each containing a separate sample, can be tested in parallel, while the mode of observation means that analysis can be combined with simultaneous measurement of conventional fluorimetric well-based assays. Here we benchmark the device against standard DEP assays, then show how such a device can be used to (a) rapidly determine the effects both of ion channel blockers on cancer cells and antibiotics on bacteria and (b) determine the properties of multiple subpopulations of cells within a well simultaneously.     

A Polymer Microfluidic Chip With Interdigitated Electrodes Arrays for Simultaneous Dielectrophoretic Manipulation and Impedimetric Detection of Microparticles

This paper presents the design, fabrication, and characterization of a polymer microfluidic biochip with integrated interdigitated electrodes arrays (IDAs) used to simultaneously separate, manipulate, and detect microparticles using dielectrophoresis (DEP) and electrochemical impedance spectroscopy (EIS) methods. The DEP response of silica microspheres has been characterized, and microspheres of different sizes (1.8 and 3.5 in diameter) have been DEP flow separated and individually trapped in different microchambers by IDAs in a single run. Simultaneously, the impedance change caused by microspheres captured on IDAs has been analyzed for quantification. High-throughput polymer microfabrication techniques such as micro injection molding were used in this work, so that the polymer microfluidic chip can be produced in a low-cost, disposable platform. This low-cost microfluidic chip provides a generic platform for developing multifunctional lab-on-a-chip devices that require the ability to handle and sense microparticles.     

A device for the fabrication of multifunctional particles from microbubble suspensions

A novel V-junction microfluidic (VJM) device has been used to generate several types of particles from the break-up of bubbles. The flow rates of selected solutions and the gas pressure were adjusted in order to successfully generate monodisperse polymer coated microbubbles, which serve as a platform for particle generation. Uniform particles in both the nano and micro size ranges with different shapes were spontaneously generated from bubbles and have the potential to be used in several advanced technological applications.     

Particle dispersion and separation resolution of pinched flow fractionation

This paper investigates a hydrodynamic particle separation technique that employs pinching of particles to a narrow microchannel. The particles are subject to a sudden expansion which results in a size-based particle separation transverse to the flow direction. The separation resolution and particle dispersion are measured using epifluorescence microscopy. The resolution and dispersion are predicted using a compact theoretical model. Devices are fabricated using conventional soft lithography of polydimethylsiloxane. The results show that the separation resolution is a function of the microchannel aspect ratio, particle size difference, and the microchannel sidewall roughness. A separation resolution as large as 3.8 is obtained in this work. This work shows that particles with diameters on the order of the sidewall roughness cannot be separated using pinched flow fractionation.     

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.     

Continuous-flow lithography for high-throughput microparticle synthesis

Precisely shaped polymeric particles and structures are widely used for applications in photonic materials, MEMS, biomaterials and self-assembly. Current approaches for particle synthesis are either batch processes or flow-through microfluidic schemes that are based on two-phase systems, limiting the throughput, shape and functionality of the particles. We report a one-phase method that combines the advantages of microscope projection photolithography and microfluidics to continuously form morphologically complex or multifunctional particles down to the colloidal length scale. Exploiting the inhibition of free-radical polymerization near PDMS surfaces, we are able to repeatedly pattern and flow rows of particles in less than 0.1 s, affording a throughput of near 100 particles per second using the simplest of device designs. Polymerization was also carried out across laminar, co-flowing streams to generate Janus particles containing different chemistries, whose relative proportions could be easily tuned. This new high-throughput technique offers unprecedented control over particle size, shape and anisotropy.     

Sheathless focusing of microbeads and blood cells based on hydrophoresis

This paper presents a microfluidic device for sheathless focusing of microbeads and blood cells based on a hydrophoretic platform comprising a V-shaped obstacle array (VOA). The VOA generates lateral pressure gradients that induce helical recirculations. Following the focusing flow particles passing through the VOA are focused in the center of the channel. In the device, the focusing pattern can be modulated by varying the gap height of the VOA. To achieve complete focusing within 4.4% coefficient of variation, the relative size differences between the gap and the particle were 3 and 4 microm for 10 and 15 microm beads, respectively. Red blood cells were used to study the hydrophoretic focusing pattern of biconcave, disk-shaped particles.     

Design and numerical investigation of a circular microchannel for particle/cell separation using dielectrophoresis

Precisely separating particles/cells with different sizes and physical properties has been an interest for point-of-care diagnostics and personalized treatment. Dielectrophoresis (DEP) is widely known as a powerful and non-invasive technique to separate particles and cells. This paper presents a comprehensive numerical investigation of particle/cell separation in circular microchannels using DEP. First, the geometrical parameters of the circular microchannel affecting DEP force are determined by performing an analytical solution. Then, by developing a solver in OpenFOAM, the effect of these parameters on particles deflection is investigated. According to the results, two different circular microchannels are presented to investigate the continuous separation of bio-particles (based on their physical properties) and polystyrene particles (based on their size). The results showed that a minimum voltage of 7, 9, and 12 V is required to achieve 100 % purity and separation efficiency for separating red blood cells from MDA-MB-231 cancer cells at the flow rate of 0.5, 1.0, and 1.5 µl/min, respectively. Also, the efficient separation of 5 and 10 µm polystyrene particles at the flow rate of 0.1 µl/min is possible only at the voltage of 9 V. The results of this numerical study can be useful for the fabrication of an optimal microdevice for the continuous DEP separation of particles and cells.     

Hydrodynamic tweezers: impact of design geometry on flow and microparticle trapping

Here we explore the role of microfabricated device geometry on frequency-dependent low Reynolds number steady streaming flow and particle trapping behavior. In our system, flow and particle trapping is induced near an obstruction or cavity located in an otherwise rectilinear oscillating flow of frequency ω and amplitude s in a fluid of kinematic viscosity ν. This work expands prior studies to characterize nine distinct obstruction/cavity geometries. The imaged microeddy flows show that the device geometry affects the eddy number, shape, structure, and strength. Comparison of measured particle trap locations with the computed eddy flow structure shows that particles trap closer to the wall than the eddy core. Trapping strength and location are controlled by the geometry and the oscillation frequency. In most cases, the trapping behavior is linearly proportional to the Stokes layer thickness, δ(AC) ~ O((ν/ω)(1/2)). We show that steady streaming in microfluidic eddies can be a flexible and versatile method for noncontact microparticle trapping, and hence we call this class of devices "hydrodynamic tweezers".