Insulator-based dielectrophoretic focusing and trapping of particles in non-Newtonian fluids

Insulator-based dielectrophoretic (iDEP) microdevices have been limited to work with Newtonian fluids. We report an experimental study of the fluid rheological effects on iDEP focusing and trapping of polystyrene particles in polyethylene oxide, xanthan gum, and polyacrylamide solutions through a constricted microchannel. Particle focusing and trapping in the mildly viscoelastic polyethylene oxide solution are slightly weaker than in the Newtonian buffer. They are, however, significantly improved in the strongly viscoelastic and shear thinning polyacrylamide solution. These observed particle focusing behaviors exhibit a similar trend with respect to electric field, consistent with a revised theoretical analysis for iDEP focusing in non-Newtonian fluids. No apparent focusing of particles is achieved in the xanthan gum solution, though the iDEP trapping can take place under a much larger electric field than the other fluids. This is attributed to the strong shear thinning-induced influences on both the electroosmotic flow and electrokinetic/dielectrophoretic motions.     

Insulator-based dielectrophoresis for the selective concentration and separation of live bacteria in water

Insulator-based dielectrophoresis (iDEP) was utilized to separate and concentrate selectively mixtures of two species of live bacteria simultaneously. Four species of bacteria were studied: the Gram-negative Escherichia coli and the Gram-positive Bacillus subtilis, B. cereus, and B. megaterium. Under an applied direct current (DC) electric field all the bacterial species exhibited negative dielectrophoretic behavior. The dielectrophoretic separations were carried out in a glass microchannel containing an array of insulating posts. The insulating posts in the microchannel produced nonuniformities in the electric field applied along the channel. Mixtures of two species of bacteria were introduced into the microchannel and the electric field was applied. The bacterial species exhibited different dielectrophoretic mobilities under the influence of the nonuniform field. From these experiments a trapping order was established with E. coli trapping at the weakest applied electric field, while the Bacillus species were trapped at different characteristic threshold fields. At stronger applied electric fields, the two different species of bacteria in the microchannel were dielectrophoretically trapped into two spatially distinct bands. The results showed that iDEP has the potential to selectively concentrate and separate different species of bacteria.     

Separation of mixtures of particles in a multipart microdevice employing insulator-based dielectrophoresis

Dielectrophoresis is the electrokinetic movement of particles due to polarization effects in the presence of non-uniform electric fields. In insulator-based dielectrophoresis (iDEP) regions of low and high electric field intensity, i.e. non-uniformity of electric field, are produced when the cross-sectional area of a microchannel is decreased by the presence of electrical insulating structures between two electrodes. This technique is increasingly being studied for the manipulation of a wide variety of particles, and novel designs are continuously developed. Despite significant advances in the area, complex mixture separation and sample fractionation continue to be the most important challenges. In this work, a microchannel design is presented for carrying out direct current (DC)-iDEP for the separation of a mixture of particles. The device comprises a main channel, two side channels and two sections of cylindrical posts with different diameters, which will generate different non-uniformities in the electric field on the main channel, designed for the discrimination and separation of particles of two different sizes. By applying an electric potential of 1000 V, a mixture of 1 and 4 μm polystyrene microspheres were dielectrophoretically separated and concentrated at the same time and then redirected to different outlets. The results obtained here demonstrate that, by carefully designing the device geometry and selecting operating conditions, effective sorting of particle mixtures can be achieved in this type of multi-section DC-iDEP devices.     

Joule heating-enabled electrothermal enrichment of nanoparticles in insulator-based dielectrophoretic microdevices

Insulator-based dielectrophoresis (iDEP) exploits the electric field gradients formed around insulating structures to manipulate particles for diverse microfluidic applications. Compared to the traditional electrode-based dielectrophoresis, iDEP microdevices have the advantages of easy fabrication, free of water electrolysis, and robust structure, etc. However, the presence of in-channel insulators may cause thermal effects because of the locally amplified Joule heating of the fluid. The resulting electrothermal flow circulations are exploited in this work to trap and concentrate nanoscale particles (of 100 nm diameter and less) in a ratchet-based iDEP microdevice. Such Joule heating-enabled electrothermal enrichment of nanoparticles are found to grow with the increase of alternating current or direct current electric field. It also becomes more effective for larger particles and in a microchannel with symmetric ratchets. Moreover, a depth-averaged numerical model is developed to understand and simulate the various parametric effects, which is found to predict the experimental observations with a good agreement.     

Quantification of pH gradients and implications in insulator-based dielectrophoresis of biomolecules

Direct current (DC) insulator-based dielectrophoretic (iDEP) microdevices have the potential to replace traditional alternating current dielectrophoretic devices for many cellular and biomolecular separation applications. The use of large DC fields suggest that electrode reactions and ion transport mechanisms can become important and impact ion distributions in the nanoliters of fluid in iDEP microchannels. This work tracked natural pH gradient formation in a 100?μm wide, 1?cm-long microchannel under applicable iDEP protein manipulation conditions. Using fluorescence microscopy with the pH-sensitive dye FITC Isomer I and the pH-insensitive dye TRITC as a reference, pH was observed to drop drastically in the microchannels within 1?min in a 3000?V/cm electric field; pH drops were observed in the range of 6-10 min within a 100?V/cm electric field and varied based on the buffer conductivity. To address concerns of dye transport impacting intensity data, electrokinetic mobilities of FITC were carefully examined and found to be (i) toward the anode and (ii) 1 to 2 orders of magnitude smaller than H? transport which is responsible for pH drops from the anode toward the cathode. COMSOL simulations of ion transport showed qualitative agreement with experimental results. The results indicate that pH changes are severe enough and rapid enough to influence the net charge of a protein or cause aggregation during iDEP experiments. The results also elucidate reasonable time periods over which the phosphate buffering capacity can counter increases in H? and OH? for unperturbed iDEP manipulations.     

Insulator-based dielectrophoretic single particle and single cancer cell trapping

Trapping of individual cells at specific locations in a microfluidic lab-on-a-chip platform is essential for single cell studies, especially those requiring individual stimulation followed by downstream analysis. To this aim, we have designed microdevices based on direct current (DC) insulator-based dielectrophoresis (iDEP) acting as individual single cell traps. We present both the design of a negative iDEP trap and a positive iDEP trap using insulating posts integrated at microchannel intersections. We obtained electric field distributions via numerical simulations adapted to the intersection and trap geometry with which we predict single particle pathlines. With polystyrene particles of 10?μm diameter, we demonstrated an effective design for a single particle trap in the case of negative dielectrophoresis. The onset trapping voltage shows an inverse relation to the buffer conductivity, thus indicating the influence of electrokinetic effects on the trapping behavior. Additionally, we demonstrated the proof-of-principle of single MCF-7 breast cancer cell trapping in a positive iDEP trap. Our single particle trapping experiments were further in very good agreement with numerical simulations. To ensure that no significant damage occurred to the cells during the experiment, we further optimized medium conditions to ensure viability of the cells for at least 1?h, more than sufficient for microfluidic trapping experiments. Our results thus indicated the successful design of DC iDEP traps, which can easily be integrated into a variety of microchip operations for single cell analysis.     

Insulator-based dielectrophoresis of microorganisms: theoretical and experimental results

Dielectrophoresis (DEP) is the motion of particles due to polarization effects in nonuniform electric fields. DEP has great potential for handling cells and is a non-destructive phenomenon. It has been utilized for different cell analysis, from viability assessments to concentration enrichment and separation. Insulator-based DEP (iDEP) provides an attractive alternative to conventional electrode-based systems; in iDEP, insulating structures are used to generate nonuniform electric fields, resulting in simpler and more robust devices. Despite the rapid development of iDEP microdevices for applications with cells, the fundamentals behind the dielectrophoretic behavior of cells has not been fully elucidated. Understanding the theory behind iDEP is necessary to continue the progress in this field. This work presents the manipulation and separation of bacterial and yeast cells with iDEP. A computational model in COMSOL Multiphysics was employed to predict the effect of direct current-iDEP on cells suspended in a microchannel containing an array of insulating structures. The model allowed predicting particle behavior, pathlines and the regions where dielectrophoretic immobilization should occur. Experimental work was performed at the same operating conditions employed with the model and results were compared, obtaining good agreement. This is the first report on the mathematical modeling of the dielectrophoretic response of yeast and bacterial cells in a DC-iDEP microdevice.     

3D Insulator‐based dielectrophoresis using DC‐biased,AC electric fields for selective bacterial trapping

Insulator‐based dielectrophoresis (iDEP) is a well‐known technique that harnesses electric fields for separating, moving, and trapping biological particle samples. Recent work has shown that utilizing DC‐biased AC electric fields can enhance the performance of iDEP devices. In this study, an iDEP device with 3D varying insulating structures analyzed in combination with DC biased AC fields is presented for the first time. Using our unique reactive ion etch lag, the mold for the 3D microfluidic chip is created with a photolithographic mask. The 3D iDEP devices, whose largest dimensions are 1 cm long, 0.18 cm wide, and 90 μm deep are then rapidly fabricated by curing a PDMS polymer in the glass mold. The 3D nature of the insulating microstructures allows for high trapping efficiency at potentials as low as 200 Vpp. In this work, separation of Escherichia coli from 1 μm beads and selective trapping of live Staphylococcus aureus cells from dead S. aureus cells is demonstrated. This is the first reported use of DC‐biased AC fields to selectively trap bacteria in 3D iDEP microfluidic device and to efficiently separate particles where selectivity of DC iDEP is limited.     

DC-biased AC-electrokinetics: a conductivity gradient driven fluid flow

This paper studies the principles of fluid flow manipulation based on DC-biased AC-electrokinetics. This method makes use of planar parallel electrodes in a microfluidic channel in contact with an electrolyte solution, with a DC biased AC electrical signal applied to the electrode pair. Due to the application of DC bias, incipient Faradaic electrolytic reactions take place resulting in an increase of the ionic content of the bulk solution. The ionic content was found to be dissimilar at the cathodic and anodic sides of the channel and a conductivity difference of approximately 10% was measured for 2 V(DC). Fluid flow is generated by the action of the DC biased AC electric signal acting on the transverse conductivity gradient generated across the microchannel. The induced flow in the form of vortex was characterized experimentally and the results substantiated theoretically. The velocity of the induced flow vortex under the employed experimental conditions was ~600 to 700 μm s(-1) which is faster than those obtained in conventional AC-electroosmosis and AC-electrothermal types of flows.     

Characterization of electrokinetic mobility of microparticles in order to improve dielectrophoretic concentration

Insulator-based dielectrophoresis (iDEP), an efficient technique with great potential for miniaturization, has been successfully applied for the manipulation of a wide variety of bioparticles. When iDEP is applied employing direct current (DC) electric fields, other electrokinetic transport mechanisms are present: electrophoresis and electroosmotic flow. In order to concentrate particles, iDEP has to overcome electrokinetics. This study presents the characterization of electrokinetic flow under the operating conditions employed with iDEP; in order to identify the optimal conditions for particle concentration employing DC-iDEP, microparticle image velocimetry (μPIV) was employed to measure the velocity of 1-μm-diameter inert polystyrene particles suspended inside a microchannel made from glass. Experiments were carried out by varying the properties of the suspending medium (conductivity from 25 to 100 μS/cm and pH from 6 to 9) and the strength of the applied electric field (50–300 V/cm); the velocities values obtained ranged from 100 to 700 μm/s. These showed that higher conductivity and lower pH values for the suspending medium produced the lowest electrokinetic flow, improving iDEP concentration of particles, which decreases voltage requirements. These ideal conditions for iDEP trapping (pH = 6 and σ _m = 100 μS/cm) were tested experimentally and with the aid of mathematical modeling. The μPIV measurements allowed obtaining values for the electrokinetic mobilities of the particles and the zeta potential of the glass surface; these values were used with a mathematical model built with COMSOL Multiphysics software in order to predict the dielectrophoretic and electrokinetic forces exerted on the particles; the modeling results confirmed the μPIV findings. Experiments with iDEP were carried out employing the same microparticles and a glass microchannel that contained an array of cylindrical insulating structures. By applying DC electric fields across the insulating structures array, it was seen that the dielectrophoretic trapping was improved when the electrokinetic force was the lowest; as predicted by μPIV measurements and the mathematical model. The results of this study provide guidelines for the selection of optimal operating conditions for improving insulator-based dielectrophoretic separations and have the potential to be extended to bioparticle applications. Figure Comparison of experimental measurements and mathematical modeling of electrokinetic and dielectrophoretic effects on microparticles

Electrothermal flow effects in insulating (electrodeless) dielectrophoresis systems

We simulate electrothermally induced flow in polymeric, insulator-based dielectrophoresis (iDEP) systems with DC-offset, AC electric fields at finite thermal Péclet number, and we identify key regimes where electrothermal (ET) effects enhance particle deflection and trapping. We study a single, two-dimensional constriction in channel depth with parametric variations in electric field, channel geometry, fluid conductivity, particle electrophoretic (EP) mobility, and channel electroosmotic (EO) mobility. We report the effects of increasing particle EP mobility, channel EO mobility, and AC and DC field magnitudes on the mean constriction temperature and particle behavior. Specifically, we quantify particle deflection and trapping, referring to the deviation of particles from their pathlines due to dielectrophoresis as they pass a constriction and the stagnation of particles due to negative dielectrophoresis near a constriction, respectively. This work includes the coupling between fluid, heat, and electromagnetic phenomena via temperature-dependent physical parameters. Results indicate that the temperature distribution depends strongly on the fluid conductivity and electric field magnitude, and particle deflection and trapping depend strongly on the channel geometry. Electrothermal (ET) effects perturb the EO flow field, creating vorticity near the channel constriction and enhancing the deflection and trapping effects. ET effects alter particle deflection and trapping responses in insulator-based dielectrophoresis devices, especially at intermediate device aspect ratios (2 ≤ r ≤ 7) in solutions of higher conductivity (σ m ≥ 1 × 10(-3)S/m). The impact of ET effects on particle deflection and trapping are diminished when particle EP mobility or channel EO mobility is high. In almost all cases, ET effects enhance negative dielectrophoretic particle deflection and trapping phenomena.     

Joule heating effects in optimized insulator‐based dielectrophoretic devices: An interplay between post geometry and temperature rise

Insulator‐based dielectrophoresis (iDEP) is the electrokinetic migration of polarized particles when subjected to a non‐uniform electric field generated by the inclusion of insulating structures between two remote electrodes. Electrode spacing is considerable in iDEP systems when compared to electrode‐based DEP systems, therefore, iDEP systems require high voltages to achieve efficient particle manipulation. A consequence of this is the temperature increase within the channel due to Joule heating effects, which, in some cases, can be detrimental when manipulating biological samples. This work presents an experimental and modeling study on the increase in temperature inside iDEP devices. For this, we studied seven distinct channel designs that mainly differ from each other in their post array characteristics: post shape, post size and spacing between posts. Experimental results obtained using a custom‐built copper Resistance Temperature Detector, based on resistance changes, show that the influence of the insulators produces a difference in temperature rise of approximately 4°C between the designs studied. Furthermore, a 3D COMSOL model is also introduced to evaluate heat generation and dissipation, which is in good agreement with the experiments. The model allowed relating the difference in average temperature for the geometries under study to the electric resistance posed by the post array in each design.     

High sensitivity three-dimensional insulator-based dielectrophoresis

Insulator-based dielectrophoresis (iDEP) is a very promising technique for sorting microparticles based on their electrical properties. The need for microfabricated electrode arrays is eliminated by using constrictions in a microchannel to induce large electric field gradients. In this work, micro-milling is used to build devices with three-dimensional features that exhibit very large constriction ratios. These three-dimensional insulator-based dielectrophoresis (3DiDEP) devices allow for trapping microparticles at average electric fields one order of magnitude lower than two-dimensional designs with the same footprint. Low voltage operation minimizes Joule heating effects that have limited previous systems, opening up the possibility for new biological applications of iDEP.     

Particle trapping in electrically driven insulator-based microfluidics: Dielectrophoresis and induced-charge electrokinetics

Electrokinetically driven insulator-based microfluidic devices represent an attractive option to manipulate particle suspensions. These devices can filtrate, concentrate, separate, or characterize micro and nanoparticles of interest. Two decades ago, inspired by electrode-based dielectrophoresis, the concept of insulator-based dielectrophoresis (iDEP) was born. In these microfluidic devices, insulating structures (i.e., posts, membranes, obstacles, or constrictions) built within the channel are used to deform the spatial distribution of an externally generated electric field. As a result, particles suspended in solution experience dielectrophoresis (DEP). Since then, it has been assumed that DEP is responsible for particle trapping in these devices, regardless of the type of voltage being applied to generate the electric field—direct current (DC) or alternating current. Recent findings challenge this assumption by demonstrating particle trapping and even particle flow reversal in devices that prevent DEP from occurring (i.e., unobstructed long straight channels stimulated with a DC voltage and featuring a uniform electric field). The theory introduced to explain those unexpected observations was then applied to conventional “DC-iDEP” devices, demonstrating better prediction accuracy than that achieved with the conventional DEP-centered theory. This contribution summarizes contributions made during the last two decades, comparing both theories to explain particle trapping and highlighting challenges to address in the near future.     

Mixing of non-Newtonian fluids in wavy serpentine microchannel using electrokinetically driven flow

A numerical investigation is performed into the mixing performance of electrokinetically driven non-Newtonian fluids in a wavy serpentine microchannel. The flow behavior of the non-Newtonian fluids is described using a power-law model. The simulations examine the effects of the flow behavior index, the wave amplitude, the wavy-wall section length, and the applied electric field strength on the mixing performance. The results show that the volumetric flow rate of shear-thinning fluids is higher than that of shear-thickening fluids, and therefore results in a poorer mixing performance. It is shown that for both types of fluid, the mixing performance can be enhanced by increasing the wave amplitude, extending the length of the wavy-wall section, and reducing the strength of the electric field. Thus, although the mixing efficiency of shear-thinning fluids is lower than that of shear-thickening fluids, the mixing performance can be improved through an appropriate specification of the flow and geometry parameters. For example, given a shear-thinning fluid with a flow behavior index of 0.8, a mixing efficiency of 87% can be obtained by specifying the wave amplitude as 0.7, the wavy-wall section length as five times the characteristic length, the nondimensional Debye-Huckel parameter as 100, and the applied electric field strength as 43.5 V/cm.     

A continuous DC-insulator dielectrophoretic sorter of microparticles

A lab-on-a-chip device is described for continuous sorting of fluorescent polystyrene microparticles utilizing direct current insulating dielectrophoresis (DC-iDEP) at lower voltages than previously reported. Particles were sorted by combining electrokinetics and dielectrophoresis in a 250 μm wide PDMS microchannel containing a rectangular insulating obstacle and four outlet channels. The DC-iDEP particle flow behaviors were investigated with 3.18, 6.20 and 10 μm fluorescent polystyrene particles which experience negative DEP forces depending on particle size, DC electric field magnitude and medium conductivity. Due to negative DEP effects, particles are deflected into different outlet streams as they pass the region of high electric field density around the obstacle. Particles suspended in dextrose added phosphate buffer saline (PBS) at conductivities ranging from 0.50 to 8.50 mS/cm at pH 7.0 were compared at 6.85 and 17.1 V/cm. Simulations of electrokinetic and dielectrophoretic forces were conducted with COMSOL Multiphysics^® to predict particle pathlines. Experimental and simulation results show the effect of medium and voltage operating conditions on particle sorting. Further, smaller particles experience smaller iDEP forces and are more susceptible to competing nonlinear electrostatic effects, whereas larger particles experience greater iDEP forces and prefer channels 1 and 2. This work demonstrates that 6.20 and 10 μm particles can be independently sorted into specific outlet streams by tuning medium conductivity even at low operating voltages. This work is an essential step forward in employing DC-iDEP for multiparticle sorting in a continuous flow, multiple outlet lab-on-a-chip device.     

Dielectrophoretic focusing of particles in a microchannel constriction using DC‐biased AC flectric fields

This paper presents a fundamental study of particle electrokinetic focusing in a single microchannel constriction. Through both experiments and simulations, we demonstrate that such dielectrophoresis‐induced particle focusing can be implemented in a much smaller magnitude of DC‐biased AC electric fields (10 kV/m in total) as compared to pure DC electric fields (up to 100 kV/m). This is attributed to the increase in the ratio of cross‐stream particle dielectrophoretic velocity to streamwise electrokinetic velocity as only the DC field component contributes to the latter. The effects of the 1 kHz frequency AC to DC electric field ratio on particle trajectories and velocity variations through the microchannel constriction are also examined, which are found to agree with the simulation results.     

Silicon insulator-based dielectrophoresis devices for minimized heating effects

Concentration of biological specimens that are extremely dilute in a solution is of paramount importance for their detection. Microfluidic chips based on insulator-based DEP (iDEP) have been used to selectively concentrate bacteria and viruses. iDEP biochips are currently fabricated with glass or polymer substrates to allow for high electric fields within the channels. Joule heating is a well-known problem in these substrates and can lead to decreased throughput and even device failure. In this work, we present, for the first time, highly efficient trapping and separation of particles in DC iDEP devices that are fabricated on silicon using a single-etch-step three-dimensional microfabrication process with greatly improved heat dissipation properties. Fabrication in silicon allows for greater heat dissipation for identical geometries and operating conditions. The 3D fabrication allows for higher performance at lower applied potentials. Thermal measurements were performed on both the presented silicon chips and previously published PDMS devices comprised of microposts. Trapping and separation of 1 and 2 μm polystyrene particles was demonstrated. These results demonstrate the feasibility of high-performance silicon iDEP devices for the next generation of sorting and concentration microsystems.     

Electrokinetic-vortex formation near a two-part cylinder with same-sign zeta potentials in a straight microchannel

Vortex formation near a two-part cylinder with zeta potentials of different values but the same sign under an external DC electric field is numerically investigated in this paper. The cylinder, inserted in a straight microchannel filled with an aqueous solution, is composed of an upstream part and a downstream part. When a DC electric field is applied in the channel, under certain conditions, the vortex will form near the cylinder due to the different velocities of electroosmotic flow generated on the cylinder surface. The numerical results reveal that the larger the velocity difference of electroosmotic flow generated on the two-part cylinder and the smaller the channel width, the more conducive to vortex formation in the channel. In addition, if the zeta potential ratios of cylinder downstream part to upstream part and channel wall to cylinder upstream part are unchanged, the DC electric field strength and the zeta potential value do not affect the pattern of vortices formed in the channel. This study provides a way for vortex formation in microchannels and has the potential application in microfluidic devices.     

Electroosmotic flow of non‐Newtonian fluids in a constriction microchannel

Insulator‐based dielectrophoresis has to date been almost entirely restricted to Newtonian fluids despite the fact that many of the chemical and biological fluids exhibit non‐Newtonian characteristics. We present herein an experimental study of the fluid rheological effects on the electroosmotic flow of four types of polymer solutions, i.e., 2000 ppm xanthan gum (XG), 5% polyvinylpyrrolidone (PVP), 3000 ppm polyethylene oxide (PEO), and 200 ppm polyacrylamide (PAA) solutions, through a constriction microchannel under DC electric fields of up to 400 V/cm. We find using particle streakline imaging that the fluid elasticity does not change significantly the electroosmotic flow pattern of weakly shear‐thinning PVP and PEO solutions from that of a Newtonian solution. In contrast, the fluid shear‐thinning causes multiple pairs of flow circulations in the weakly elastic XG solution, leading to a central jet with a significantly enhanced speed from before to after the channel constriction. These flow vortices are, however, suppressed in the strongly viscoelastic and shear‐thinning PAA solution.