Insulator-based Dielectrophoresis Research Articles

Enhanced horizontal alignment of InGaN/GaN nanorod LEDs via insulator-based dielectrophoresis

GaN-based nanorod light emitting diodes (LEDs) are key to the development of high brightness, small pixel size, and long lifetime for high-resolution display applications. To manufacture individual nanorod LEDs as compact light sources, it is necessary to separate the nanorod LEDs from the substrate and transfer them to the electrode. Dielectrophoresis (DEP) is a highly suitable technique for transferring the individual nanorod LEDs. However, there are still challenges in achieving a high alignment yield. In this work, nanorod LEDs are successfully fabricated and separated to enhance the horizontal alignment in interdigitated electrodes via insulator-based DEP (iDEP). In iDEP, a structure is formed by inserting an insulating layer between the electrodes. This structure manipulates the electric field generated around the electrodes due to polarization by the insulating layer. This results in a relatively uniform electric field distribution on the surface of the insulating layer, thereby enhancing the horizontal alignment of the nanorod LEDs. Herein, the iDEP structure achieves significant nanorod LED alignment yield of up to 91.9 %, compared to 45.5 % for the electrode-based DEP (eDEP) technique. Furthermore, the use of a patterned insulator makes it possible to restrict 1 or 2 aligned nanorod LEDs within a specific pixel region, which is suitable for high-resolution display technologies based on nanorod LEDs.

Insulator-based dielectrophoresis-assisted separation of insulin secretory vesicles.

Organelle heterogeneity and inter-organelle contacts within a single cell contribute to the limited sensitivity of current organelle separation techniques, thus hindering organelle subpopulation characterization. Here, we use direct current insulator-based dielectrophoresis (DC-iDEP) as an unbiased separation method and demonstrate its capability by identifying distinct distribution patterns of insulin vesicles from INS-1E insulinoma cells. A multiple voltage DC-iDEP strategy with increased range and sensitivity has been applied, and a differentiation factor (ratio of electrokinetic to dielectrophoretic mobility) has been used to characterize features of insulin vesicle distribution patterns. We observed a significant difference in the distribution pattern of insulin vesicles isolated from glucose-stimulated cells relative to unstimulated cells, in accordance with maturation of vesicles upon glucose stimulation. We interpret the difference in distribution pattern to be indicative of high-resolution separation of vesicle subpopulations. DC-iDEP provides a path for future characterization of subtle biochemical differences of organelle subpopulations within any biological system.

Charge-Based Separation of Microparticles Using AC Insulator-Based Dielectrophoresis.

Surface charge is an important property of particles. It has been utilized to separate particles in microfluidic devices, where dielectrophoresis (DEP) is often the driving force. However, current DEP-based particle separations based on the charge differences work only for particles of similar sizes. They become less effective and may even fail for a mixture of particles differing in both charge and size. We demonstrate that our recently developed AC insulator-based dielectrophoresis (AC iDEP) technique can direct microparticles toward charge-dependent equilibrium positions in a ratchet microchannel. Such charge-based particle separation is controlled by the imposed AC voltage frequency and amplitude but is nearly unaffected by the size of either type of particle in the mixture except for the time required to achieve an effective separation. This AC iDEP technique may potentially be used to focus and separate submicron or even nanoparticles because of its virtually "infinite" channel length.

Investigating the use of direct current insulator-based dielectrophoresis to study insulin vesicle subpopulations

Investigating the use of direct current insulator-based dielectrophoresis to study insulin vesicle subpopulations

On the behavior of sub-micrometer polystyrene particles subjected to AC insulator-based dielectrophoresis.

Polymer beads, especially polystyrene particles, have been extensively used as model species in insulator-based dielectrophoresis (iDEP) studies. Their use in alternating current iDEP (AC-iDEP) is less explored; however, an assessment in the low-frequency regime (≤10kHz) allows to link surface conduction effects with the surface properties of polymer particles. Here, we provide a case study for various experimental conditions assessing sub-micrometer polystyrene particles with AC-iDEP and link to accepted surface conduction theory to predict and experimentally verify the observed AC-iDEP trapping behavior based on apparent zeta potential and solution conductivity. We find excellent agreement with the theoretical predictions, but also the occurrence of concentration polarization electroosmotic flow under the studied conditions, which have the potential to confound acting dielectrophoresis conditions. Furthermore, we study a case relevant to the assessment of microplastics in human and animal body fluids by mimicking the protein adsorption of high abundant proteins in blood by coating polystyrene beads with bovine serum albumin, a highly abundant protein in blood. Theoretical predictions and experimental observations confirm a difference in observed AC-iDEP behavior between coated and non-coated particles, which might be exploited for future studies of microplastics in blood to assess their exposure to humans and animals.

High-Throughput Continuous Free-Flow Dielectrophoretic Trapping of Micron-Scale Particles and Cells in Paper Using Localized Nonuniform Pore-Scale-Generated Paper-Based Electric Field Gradients.

Dielectrophoresis (DEP) utilizes a spatially varying nonuniform electrical field to induce forces on suspended polarizable soft matter including particles and cells. Such nonuniformities are conventionally created using 2D or 3D micrometer-scale electrode arrays. Alternatively, insulator-based dielectrophoresis (iDEP) uses small micrometer-scale insulating structures to spatially distort and generate regions of localized field gradients to selectively trap, isolate, and concentrate bioparticles, including bacteria, viruses, red blood cells, and cancer cells from a suspending electrolyte solution. Despite significant advances in the microfabrication technology, the commercial adoption of DEP devices for soft matter manipulation remains elusive. One reason for low market penetration is a lack of low-cost and scalable fabrication methods to quickly microfabricate field-deforming structures to generate localized DEP-inducing electric field gradients. We propose here that paper-based devices can offer a low-cost and easy-to-use alternative to traditional iDEP devices. In this article, we demonstrate for the first time the ability to perform iDEP-style particle trapping using the naturally occurring micrometer-scale insulating porous structures of paper. In particular, we use polymeric laminated nonwoven fiberglass paper channels as a source of insulating structures for iDEP. We apply a flow of polarizable microparticles directly within the nonwoven channel and simultaneously drop an electric field perpendicular to the flow direction to induce DEP. We show the ability to readily trap and concentrate particles in paper by DEP with an applied voltage as low as 2 V using two different flow mechanisms: a constant fluid flow rate using an external pump and passive fluid flow by capillary wicking. Using a combination of micro computed tomography and finite element analysis, we then present a computational model to probe the microscale DEP force formation dynamics within the paper structure. This new paper-based iDEP platform enables the development of robust, low-cost, and portable next-generation iDEP systems for a wide variety of sample purification and liquid handling applications.

Fluid Elasticity-Enhanced Insulator-Based Dielectrophoresis for Sheath-Free Particle Focusing in Very Dilute Polymer Solutions.

Focusing particles into a narrow stream is usually a necessary step in microfluidic flow cytometry and particle sorting. We demonstrate that the addition of a small amount of poly(ethylene oxide) (PEO) polymer into a buffer solution can reduce by almost 1 order of magnitude the threshold DC electric field for single-line dielectrophoretic focusing of particles in a constricted microchannel. The particle focusing effectiveness of this fluid elasticity-enhanced insulator-based dielectrophoresis (E-iDEP) in very dilute PEO solutions gets enhanced with the increase of the PEO molecular weight and particle size. These two trends are consistent with a theoretical analysis that accounts for the fluid elasticity effects on the electrokinetic and dielectrophoretic particle motions. Surprisingly, the particle-focusing effectiveness of E-iDEP is observed to first increase and then decrease with an increase in the PEO concentration.

Numerical modeling reveals improved organelle separation for dielectrophoretic ratchet migration.

Organelle size varies with normal and abnormal cell function. Thus, size-based particle separation techniques are key to assessing the properties of organelle subpopulations differing in size. Recently, insulator-based dielectrophoresis (iDEP) has gained significant interest as a technique to manipulate sub-micrometer-sized particles enabling the assessment of organelle subpopulations. Based on iDEP, we recently reported a ratchet device that successfully demonstrated size-based particle fractionation in combination with continuous flow sample injection. Here, we used a numerical model to optimize the performance with flow rates a factor of three higher than previously and increased the channel volume to improve throughput. We evaluated the amplitude and duration of applied low-frequency DC-biased AC potentials improving separation efficiency. A separation efficiency of nearly 0.99 was achieved with the optimization of key parameters-improved from 0.80 in previous studies (Ortiz et al. Electrophoresis, 2022;43;1283-1296)-demonstrating that fine-tuning the periodical driving forces initiating the ratchet migration under continuous flow conditions can significantly improve the fractionation of organelles of different sizes.

A numerical study on microfluidic devices to maintain the concentration and purity of dielectrophoresis-induced separated fractions of analyte.

Determining the physical and chemical properties of biologically important particles such as cells, organelles, viruses, exosomes, complexes, nucleotides, and proteins is needed to understand their function. These properties are determined with common analytical tools (mass spectrometry, cryo-EM, NMR, various spectroscopies, nucleotide sequencing, etc.) whose function can be improved when samples are pure and concentrated. Separations science plays acentral role in conditioning samples, ranging from low-resolution benchtop operations like precipitations or extractions to higher-resolution chromatography and electrophoresis. In the last two decades, gradient insulator-based dielectrophoresis (g-iDEP) has emerged as a high-resolution separation technique capable of highly selective enrichment of cells, viruses, exosomes, and proteins. Specific evidence has been shown that pure homogeneous and concentrated fractions of cells and exosomes can be generated from complex mixtures. However, recovering those fractions for analysis has not been developed, limiting the technique to an analytical rather than a preparative one. Here, a finite element analysis was undertaken to identify geometries and operational parameters to efficiently remove the enriched fraction while retaining maximum concentration and providing total mass transfer. Geometric factors (e.g., side channel width and distance from the gradient-inducing gap) were studied, along with the addition of a second inlet side channel. Two flow-generating mechanisms-electroosmosis and hydrostatic pressure-were evaluated for semi-optimized device designs, including a comparison of the one- and two-inlet designs. Simulations indicate effectively one hundred percent mass transfer and a concentration increase by an order of magnitude for several device configurations and operational parameters.

Numerical study of a double-stair-shaped dielectrophoresis channel for continuous on-chip cell separation and lysis using finite element method

Prognostication of numerous chronic diseases are in need of identifying circulating tumor cells (CTCs), afterwards, separating and reviving contaminated samples are required. Conventional methods of separating blood cells, namely cytometry or magnetically activated cell sorting, in many cases lose their functionality, or efficiency under different conditions. Hence microfluidic methods of separation have been implemented. Herein, an innovative integrated double stair-shaped microchannel is designed and optimized, capable of ‘separation’, and ‘chemical lysis’ simultaneously in which the lysis reagent concentration can be controlled to tune the lysis intensity. The method of insulator-based dielectrophoresis (iDEP), which is the main physics in this device, is utilized yielding maximum separation. Pivotal features of the applied voltage, the voltage difference, the angles and the number of stairs, and the width of the throat in the microchannel have been numerically explored in order to optimize the channel in terms of separation and the lysis buffer concentration. The overall state of optimum case for the voltage difference (ΔV) of 10 owns the following features: the number of stairs is 2, the angle of stairs is 110°, the width of throat is 140 μm, and the inlet voltages are 30 V and 40 V. Also, the overall state of optimum cases for delta possess the following features: the number of stairs is 2, the angle of stairs is 110°, the width of throat is 140 μm, and the inlet voltages are 30 V and 35 V.

Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coli.

Lipopolysaccharide (LPS) is the unique feature that composes the outer leaflet of the Gram-negative bacterial cell envelope. Variations in LPS structures affect a number of physiological processes, including outer membrane permeability, antimicrobial resistance, recognition by the host immune system, biofilm formation, and interbacterial competition. Rapid characterization of LPS properties is crucial for studying the relationship between these LPS structural changes and bacterial physiology. However, current assessments of LPS structures require LPS extraction and purification followed by cumbersome proteomic analysis. This paper demonstrates one of the first high-throughput and non-invasive strategies to directly distinguish Escherichia coli with different LPS structures. Using a combination of three-dimensional insulator-based dielectrophoresis (3DiDEP) and cell tracking in a linear electrokinetics assay, we elucidate the effect of structural changes in E. coli LPS oligosaccharides on electrokinetic mobility and polarizability. We show that our platform is sufficiently sensitive to detect LPS structural variations at the molecular level. To correlate electrokinetic properties of LPS with the outer membrane permeability, we further examined effects of LPS structural variations on bacterial susceptibility to colistin, an antibiotic known to disrupt the outer membrane by targeting LPS. Our results suggest that microfluidic electrokinetic platforms employing 3DiDEP can be a useful tool for isolating and selecting bacteria based on their LPS glycoforms. Future iterations of these platforms could be leveraged for rapid profiling of pathogens based on their surface LPS structural identity.

Separation of bacteria smaller than 4µm from other blood components using insulator-based dielectrophoresis: numerical simulation approach.

Bloodstream infection (BSI) is a life-threatening infection that causes more than 80,000 deaths and more than 500,000 infections annually in North America. The rapid diagnosis of infection reduces BSI mortality. We proposed bacterial enrichment and separation approach in the current work that may reduce culturing time and accelerate the diagnosis of infection. Over the last two decades, multiple separation methods have been developed, and among these methods, insulator-based dielectrophoresis (iDEP) is considered a powerful technique for separating biological particles. Bacterial separation in the blood is challenging due to the presence of other blood cells, such as white blood cells, red blood cells, and platelets. In the present study, a model is presented which is capable of blood cells separation and directing each cell to a specific outlet using continuous flows of particles with sizes larger than 8µm, 8-4µm, and smaller than 4µm. Compared to other methods, such as filtration, the main advantage of this model is that particles larger than 8µm are separated from the flow before other particles, which prevents the accumulation of particles in the channel. The outcomes of simulations demonstrated that the factors such as applied voltage and channel dimensions significantly affect the separation efficiency. If these values are properly selected (for example voltage of 70V that was causing an electric field of 200V/cm), the proposed model can completely (100%) separate particles larger than 8µm and smaller than 4µm (8-4µm particles separation efficiency is 95%).

Selective Trapping of Living Cells into Micropores within a Track-etched Membrane by Dielectrophoresis

In this study, an insulator-based dielectrophoresis device using a track-etched membrane was developed for selective trapping of living yeast cells. A high electric field was formed in cylindrical pores of 8-µm diameter within the insulating membrane between two transparent plate electrodes. At a frequency of 20 MHz, only a living cell could be drawn into the pore and trapped strongly by dielectrophoresis. For non-spherical cells with a diameter and length of 5 µm and 14 µm, respectively, it was found that a cell longer than the pore diameter was drawn into the pores with electro-orientation if the minor axis of the cell was shorter.

Parallelized continuous flow dielectrophoretic separation ofDNA.

Numerous microfluidic separation applications have been shown in the past years providing a fast analysis of biological samples like DNA or proteins. Microfluidic separation based on dielectrophoresis (DEP), that is the migration of a polarizable object in an inhomogeneous electric field, provides numerous advantages. However, the main drawback of DEP separation devices is that they are not sufficient for large-scale sample purification due to the lack of high sample throughput. In this work, we present for the first time a microfluidic device with two parallelized dielectrophoretic separations of (biological) samples smaller than 1µm. The separation is carried out by means of insulator-based DEP, that is an insulating ridge reduced the flow through height and thus created a nanoslit at which the selective DEP forces occur. The device consists of a cross injector, two parallel operation regions and separate harvesting reservoirs where the samples are collected. Each DEP operation region contains an insulating ridge. We successfully demonstrate the separation of 100 and 40nm beads and 10 and 5kbp DNA with a separation purity of more than 80%. This states the proof-of-concept for up-scaling of dielectrophoretic separation by parallelization. As the present technique is virtually label-free, it offers a fast purification, for example in the production of gene vaccines.

Electrokinetic separation of cfDNA in insulator-based dielectrophoresis systems: a linear model of cfDNA and investigation of effective parameters

In this study, a comprehensive numerical simulation was done to investigate the electrokinetic translocation of cfDNA molecule as well as the possibility of its detection and separation in insulator based dielectrophoresis (iDEP) systems. Modeling was done for the first time by solving the Poisson equation for the electrical potential, Naiver–Stokes (NS) equation for the fluid flow and energy equation for the heat transfer in the system and considering a coarse-grained bead-spring model to describe the conformational and geometrical changes of cfDNA molecule. The effect of the geometrical parameters of the system, the initial orientation of the molecule, electrical conductivity of the solution and zeta potential of the wall was investigated on the translocation and the minimum voltage required for cfDNA trapping. When the ratio of the inlet height to the constriction zone height is large enough, cfDNA molecules cannot pass through the nanopore and trap in the constriction zone. Also, it was found that the electrical conductivity of the solution is a limiting parameter to directly isolate cfDNA from pure plasma without dilution due to significant increase in the temperature of the system. Our results demonstrate the enormous potential of iDEP systems for rapid detection of cfDNA from diluted plasma under special electrical potential and geometrical parameters of the iDEP systems.

Continuous organelle separation in an insulator-based dielectrophoretic device.

Heterogeneity in organelle size has been associated with devastating human maladies such as neurodegenerative diseases or cancer. Therefore, assessing the size-based subpopulation of organelles is imperative to understand the biomolecular foundations of these diseases. Here, we demonstrated a ratchet migration mechanism using insulator-based dielectrophoresis in conjunction with a continuous flow component that allows the size-based separation of submicrometer particles. The ratchet mechanism was realized in a microfluidic device exhibiting an array of insulating posts, tailoring electrokinetic and dielectrophoretic transport. A numerical model was developed to elucidate the particle migration and the size-based separation in various conditions. Experimentally, the size-based separation of a mixture of polystyrene beads (0.28 and 0.87 m) was accomplished demonstrating good agreement with the numerical model. Furthermore, the size-based separation of mitochondria was investigated using a mitochondria mixture isolated from HepG2 cells and HepG2 cells carrying the gene Mfn-1 knocked out, indicating distinct size-related migration behavior. With the presented continuous flow separation device, larger amounts of fractionated organelles can be collected in the future allowing access to the biomolecular signature of mitochondria subpopulations differing in size.

Individual Microparticle Manipulation Using Combined Electroosmosis and Dielectrophoresis through a Si3N4 Film with a Single Micropore.

Porous dielectric membranes that perform insulator-based dielectrophoresis or electroosmotic pumping are commonly used in microchip technologies. However, there are few fundamental studies on the electrokinetic flow patterns of single microparticles around a single micropore in a thin dielectric film. Such a study would provide fundamental insights into the electrokinetic phenomena around a micropore, with practical applications regarding the manipulation of single cells and microparticles by focused electric fields. We have fabricated a device around a silicon nitride film with a single micropore (2–4 µm in diameter) which has the ability to locally focus electric fields on the micropore. Single microscale polystyrene beads were used to study the electrokinetic flow patterns. A mathematical model was developed to support the experimental study and evaluate the electric field distribution, fluid motion, and bead trajectories. Good agreement was found between the mathematic model and the experimental data. We show that the combination of electroosmotic flow and dielectrophoretic force induced by direct current through a single micropore can be used to trap, agglomerate, and repel microparticles around a single micropore without an external pump. The scale of our system is practically relevant for the manipulation of single mammalian cells, and we anticipate that our single-micropore approach will be directly employable in applications ranging from fundamental single cell analyses to high-precision single cell electroporation or cell fusion.

Fluid rheological effects on streaming dielectrophoresis in a post-array microchannel.

Recent studies have demonstrated the strong influences of fluid rheological properties on insulator-based dielectrophoresis (iDEP) in single-constriction microchannels. However, it is yet to be understood how iDEP in non-Newtonian fluids depends on the geometry of insulating structures. We report in this work an experimental study of fluid rheological effects on streaming DEP in a post-array microchannel that presents multiple contractions and expansions. The iDEP focusing and trapping of particles in a viscoelastic polyethylene oxide solution are comparable to those in a Newtonian buffer, which is consistent with the observations in a single-constriction microchannel. Similarly, the insignificant iDEP effects in a shear-thinning xanthan gum solution also agree with those in the single-constriction channel except that gel-like structures are observed to only form in the post-array microchannel under large DC electric fields. In contrast, the iDEP effects in both viscoelastic and shear-thinning polyacrylamide solution are significantly weaker than in the single-constriction channel. Moreover, instabilities occur in the electroosmotic flow and appear to be only dependent on the DC electric field. These phenomena may be associated with the dynamics of polymers as they are electrokinetically advected around and through the posts.

The latest advances on nonlinear insulator-based electrokinetic microsystems under direct current and low-frequency alternating current fields: a review

This review article presents an overview of the evolution of the field of insulator-based dielectrophoresis (iDEP); in particular, it focuses on insulator-based electrokinetic (iEK) systems stimulated with direct current and low-frequency(< 1kHz) AC electric fields. The article covers the surge of iDEP as a research field where many different device designs were developed, from microchannels with arrays of insulating posts to devices with curved walls and nano- and micropipettes. All of these systems allowed for the manipulation and separation of a wide array of particles, ranging from macromolecules to microorganisms, including clinical and biomedical applications. Recent experimental reports, supported by important theoretical studies in the field of physics and colloids, brought attention to the effects of electrophoresis of the second kind in these systems. These recent findings suggest that DEP is not the main force behind particle trapping, as it was believed for the last two decades. This new research suggests that particle trapping, under DC and low-frequency AC potentials, mainly results from a balance between electroosmotic and electrophoretic effects (linear and nonlinear); although DEP is present in these systems, it is not a dominant force. Considering these recent studies, it is proposed to rename this field from DC-iDEP to DC-iEK (and low-frequency AC-iDEP to low-frequency AC-iEK). Whereas much research is still needed, this is an exciting time in the field of microscale EK systems, as these new findings seem to explain the challenges with modeling particle migration and trapping in iEK devices, and provide perhaps a better understanding of the mechanisms behind particle trapping.

Interplay of induced charge electroosmosis and electrothermal flow in insulator-based dielectrophoresis

A depth-averaged numerical model is developed to understand the experimentally observed interplay of induced charge electroosmosis and electrothermal flow in an insulator-based dielectrophoresis microdevice. The experimentally measured nonlinear fluid velocity matches asymptotically the predicted velocity of the electroosmotic flow in a low-concentration buffer and that of electrothermal flow in a high-concentration buffer. This agreement is consistent with a scaling analysis.