Dielectrophoresis Research Articles

Electrochemical Impedance Spectroscopy in the Determination of the Dielectric Properties of Tau-441 Protein for Dielectrophoresis Response Prediction.

This study employs electrochemical impedance spectroscopy (EIS) to probe the behavior of Tau-441 protein, a key component implicated in Alzheimer's disease. Through meticulous experimentation and analysis, the impedance of Tau-441 protein suspension revealed a conductivity peak value of 1.02 S/m. The study demonstrates a high level of specificity and selectivity, particularly within the challenging nanomolar concentration range. Additionally, the EIS method enabled the prediction of Tau-441 protein's dielectrophoresis (DEP) response and the determination of the associated frequency range of 1 kHz to 1 MHz. These findings contribute to advancing our understanding of the molecular intricacies surrounding Tau-441 and hold promise for unraveling implications related to Alzheimer's disease. This study establishes a robust foundation for future research on neurodegenerative disease and biosciences, offering valuable insights into the electrochemical dynamics of Tau-441 protein.

Topology optimization of the electrodes in dielectrophoresis-based devices

This paper aims for developing topology optimization methodology to design the shape of electrodes in Dielectrophoresis (DEP)-based devices. The DEP force is due to a non-uniform electric field induced by applied voltages to the electrodes. Shape of the electrodes has the principal effect on the direction and magnitude of the DEP force. In medical therapy microfluidic devices, DEP force is used for cell sorting and cell separation. While the direction and magnitude of the DEP force are desired to be determined and maximized respectively, the magnitude of the electric field should be minimized to avoid damaging cells. Approaching these goals is counter intuitive where the existing electrode designs are basic. Therefore, a detailed finite element model (FEM) is developed for DEP force and electric field to formulate an optimization problem to maximize the DEP force in a particular direction while there is a constraint on electric field's magnitude. Using the developed FEM, explicit formulations for sensitivity analysis are derived to implement a gradient-based topology optimization. The performance of developed methodology is assessed numerically to determine the direction of the DEP force and constraining the electric field and experimentally in a practical case study of particle trapping in a microfluidic channel.

Integration Of Acoustophoresis and Dielectrophoresis on A Single Chip Platform: A Numerical Study

Passive or active manipulation forces would be needed for the microfluidic system for chemical and biological analysis in order to regulate, trap, separate, sort, and discriminate between particles and cells. The primary goals of passive manipulation are consistency and repeatability to attain high levels of control with exact trajectories. In the meantime, by introducing external forces like hydrodynamic, dielectrophoretic, magnetophoretic, acoustophoretic, and optical tweezing, the active manipulations allow to control particle displacement in a highly predictable and consistent fashion. These methods are much more promising for the development of a small and compact biomedical diagnostic rapid test. Since most biological particles are suspended in different biological fluids like blood and urine, dielectrophoresis (DEP) and acoustophoresis (ACP) have been demonstrated to be promising among these external forces because of their ability to apply forces on the particles in a liquid environment. Additionally, both techniques are fast, inexpensive to fabricate, label-free, and incredibly selective. In this study, we introduce a novel method that combines these two forces into a single chip, improving the separation process for ACP and DEP based on the intrinsic dielectric and acoustic properties of the particles, respectively. It is anticipated that this research would shed light on why particular manipulative factors predominate more or less in particular situations.

Dielectrophoretic motion of a red blood cell in a microfluidic environment: Insights from numerical simulations

This research delves into the dielectrophoresis (DEP) behavior of a biological cell within a sinusoidal-shaped microchannel utilizing the Maxwell stress tensor (MST) theory. A red blood cell (RBC), immersed in a viscoelastic fluid, is studied considering the Oldroyd-B model. The study aims to fill a gap in the literature by examining the DEP characteristics of RBC in a realistic geometric configuration and fluid environment, bridging the divide between theoretical modeling and practical application. This work uniquely explores the DEP behavior of an RBC within a sinusoidal microchannel in the presence of a viscoelastic flow regime, which simulates plasma properties, marking a novel contribution to the field. The two-dimensional numerical model incorporates the finite element method to accurately simulate the DEP effect and describe the behavior of the viscoelastic fluid. Validation results confirm the accuracy of the MST model. Crucially, numerical findings highlight the strong dependence of DEP force on electric potential and fluid permittivity. As a consequence of their heightened levels, there is an associated increase in both the DEP force and velocity. While the augmentation of fluid viscosity merely results in a deceleration of DEP velocity. The study provides valuable insights into the interplay between physical parameters and particle behavior, paving the way for advancements in microfluidic particle manipulation techniques.

Rapid and Sensitive Detection by Combining Electric Field Effects and Surface Plasmon Resonance: A Theoretical Study.

Surface plasmon resonance (SPR) has been extensively employed in biological sensing, environmental detection, as well as chemical industry. Nevertheless, the performance possessed by conventional surface plasmon resonance (SPR) biosensors can be further limited by the transport of analyte molecules to the sensing surface, noteworthily when small molecules or low levels of substances are being detected. In this study, a rapid and highly sensitive SPR biosensor is introduced to enhance the ability of the target analytes' collection by integrating AC electroosmosis (ACEO) and dielectrophoresis (DEP). Both the above-mentioned phenomena principally arise from the generation of the AC electric fields. This generation can be tailored by shaping the interdigitated electrodes (IDEs) that also serve as the SPR biomarker sensing area. The effects exerted by different parameters (e.g., the frequency and voltage of the AC electric field as well as microelectrode structures) are considered in the iSPR (interdigitated SPR) biosensor operation, and the iSPR biosensors are optimized with the sensitivity. The results of this study confirm that the iSPR can efficiently concentrate small molecules into the SPR sensing area, such that SPR reactions achieve an order of magnitude increase, and the detection time is shortened. The rapid and sensitive sensor takes on critical significance in the development of on-site diagnostics in a wide variety of human and animal health applications.

Dielectrophoresis: Measurement technologies and auxiliary sensing applications.

Dielectrophoresis (DEP), which arises from the interaction between dielectric particles and an aqueous solution in a nonuniform electric field, contributes to the manipulation of nano and microparticles in many fields, including colloid physics, analytical chemistry, molecular biology, clinical medicine, and pharmaceutics. The measurement of the DEP force could provide a more complete solution for verifying current classical DEP theories. This review reports various imaging, fluidic, optical, and mechanical approaches for measuring the DEP forces at different amplitudes and frequencies. The integration of DEP technology into sensors enables fast response, high sensitivity, precise discrimination, and label-free detection of proteins, bacteria, colloidal particles, and cells. Therefore, this review provides an in-depth overview of DEP-based fabrication and measurements. Depending on the measurement requirements, DEP manipulation can be classified into assistance and integration approaches to improve sensor performance. To this end, an overview is dedicated to developing the concept of trapping-on-sensing, improving its structure and performance, and realizing fully DEP-assisted lab-on-a-chip systems.

Electrical signature of heterogeneous human mesenchymal stem cells.

Human mesenchymal stem cells (hMSCs) have gained traction in transplantation therapy due to their immunomodulatory, paracrine, immune-evasive, and multipotent differentiation potential. The inherent heterogeneity of hMSCs poses a challenge for therapeutic treatments and necessitates the identification of robust biomarkers to ensure reproducibility in both in vivo and in vitro experiments. In this study, we utilized dielectrophoresis (DEP), a label-free electrokinetic phenomenon, to investigate the heterogeneity of hMSCs derived from bone marrow (BM) and adipose tissue (AD). The electrical properties of BM-hMSCs were compared to homogeneous mouse fibroblasts (NIH-3T3), human fibroblasts (WS1), and human embryonic kidney cells (HEK-293). The DEP profile of BM-hMSCs differed most from HEK-293 cells. We compared the DEP profiles of BM-hMSCs and AD-hMSCs and found that they have similar membrane capacitances, differing cytoplasm conductivity, and transient slopes. Inducing both populations to differentiate into adipocyte and osteoblast cells revealed that they behave differently in response to differentiation-inducing cytokines. Histology and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analyses of the differentiation-related genes revealed differences in heterogeneity between BM-hMSCs and AD-hMSCs. The differentiation profiles correlate well with the DEP profiles developed and indicate differences in the heterogeneity of BM-hMSCs and AD-hMSCs. Our results demonstrate that using DEP, membrane capacitance, cytoplasm conductivity, and transient slope can uniquely characterize the inherent heterogeneity of hMSCs to guide robust and reproducible stem cell transplantation therapies.

A Label-Free Droplet Sorting Platform Integrating Dielectrophoretic Separation for Estimating Bacterial Antimicrobial Resistance.

Antimicrobial resistance (AMR) has become a crucial global health issue. Antibiotic-resistant bacteria can survive after antibiotic treatments, lowering drug efficacy and increasing lethal risks. A microfluidic water-in-oil emulsion droplet system can entrap microorganisms and antibiotics within the tiny bioreactor, separate from the surroundings, enabling independent assays that can be performed in a high-throughput manner. This study presents the development of a label-free dielectrophoresis (DEP)-based microfluidic platform to sort droplets that co-encapsulate Escherichia coli (E. coli) and ampicillin (Amp) and droplets that co-encapsulate Amp-resistant (AmpR) E. coli with Amp only based on the conductivity-dependent DEP force (FDEP) without the assistance of optical analyses. The 9.4% low conductivity (LC) Luria-Bertani (LB) broth diluted with 170 mM mannitol can maintain E. coli and AmpR E. coli growth for 3 h and allow Amp to kill almost all E. coli, which can significantly increase the LCLB conductivity by about 100 μS/cm. Therefore, the AmpR E. coli/9.4%LCLB/Amp where no cells are killed and the E. coli/9.4%LCLB/Amp-containing droplets where most of the cells are killed can be sorted based on this conductivity difference at an applied electric field of 2 MHz and 100 Vpp that generates positive FDEP. Moreover, the sorting ratio significantly decreased to about 50% when the population of AmpR E. coli was equal to or higher than 50% in droplets. The conductivity-dependent DEP-based sorting platform exhibits promising potential to probe the ratio of AmpR E. coli in an unknown bacterial sample by using the sorting ratio as an index.

Computational Studies Using COMSOL Multiphysics on Dielectrophoretic Characterization and Separation of Mesenchymal Stem Cells Undergoing Tenogenic Differentiation

Stem cells have unique self-renewal and differentiation capacities, which are advantageous for regenerative medicine and tissue engineering applications. Stem cells can recreate cells needed to repair injured tissues and organs in the body, for example, by regenerating connective tissues, which provide support, protection, and a structural framework for various organs and other tissues. However, a significant limitation is the susceptibility of tendons to injury with long-term loss of function. Mesenchymal Stem Cell (MSC) therapies are promising for healing tendon injuries and tears but controlling stem cell differentiation and generating homogeneous tenogenic stem cell populations remains challenging. Therefore, separating differentiating and non-differentiating MSCs to collect homogenous cells is critical for further tissue repair after transplantation. To substantiate our hypothesis that differentiating and non-differentiating stem cells have unique dielectric properties, we focused on characterizing-obtaining dielectric signatures for differentiating tenogenic and non-tenogenic MSCs on the third day of differentiation using dielectrophoresis (DEP) focused on crossover frequencies [1, 2]. The estimated values for membrane capacitance, conductivity, and permittivity after 3-day treatment of undifferentiated cells differentiating into tenogenic MSCs are 2.46±0.1 pF, 0.82±0.01 S/m, and 1.97±0.05, respectively, and cytoplasm conductivity 0.82±0.02 S/m. The treated stem cells undergo differentiation, but the untreated cells don’t. The results showed a significant difference between the 3-day treatment and no-treatment group. The estimated properties from the dielectrophoretic characterization studies are crucial in designing a DEP-based enrichment microdevice to collect homogeneous differentiated stem cell populations for tendon repair. Using particle tracing, creeping flow (transport of diluted species model), and electric current physics in COMSOL Multiphysics simulation software, we designed a microdevice that achieves 100% separation of untreated and treated mesenchymal stem cells undergoing differentiation towards tendon. Applying dielectrophoresis to the microdevice architecture demonstrated high selectivity and yield, processing one million treated and untreated stem cells in 6.4 and 27.8 hours, respectively.

A Novel Microfluidic Strategy for Efficient Exosome Separation via Thermally Oxidized Non-Uniform Deterministic Lateral Displacement (DLD) Arrays and Dielectrophoresis (DEP) Synergy.

Exosomes, with diameters ranging from 30 to 150 nm, are saucer-shaped extracellular vesicles (EVs) secreted by various type of human cells. They are present in virtually all bodily fluids. Owing to their abundant nucleic acid and protein content, exosomes have emerged as promising biomarkers for noninvasive molecular diagnostics. However, the need for exosome separation purification presents tremendous technical challenges due to their minuscule size. In recent years, microfluidic technology has garnered substantial interest as a promising alternative capable of excellent separation performance, reduced reagent consumption, and lower overall device and operation costs. In this context, we hereby propose a novel microfluidic strategy based on thermally oxidized deterministic lateral displacement (DLD) arrays with tapered shapes to enhance separation performance. We have achieved more than 90% purity in both polystyrene nanoparticle and exosome experiments. The use of thermal oxidation also significantly reduces fabrication complexity by avoiding the use of high-precision lithography. Furthermore, in a simulation model, we attempt to integrate the use of dielectrophoresis (DEP) to overcome the size-based nature of DLD and distinguish particles that are close in size but differ in biochemical compositions (e.g., lipoproteins, exomeres, retroviruses). We believe the proposed strategy heralds a versatile and innovative platform poised to enhance exosome analysis across a spectrum of biochemical applications.

Approaches for attaining clean bacterial fractions from complex environmental samples

Marine bacteria have been targeted by industry and pharmaceutics as genetic resources for highly active enzymes or novel lead compounds. Although numerous techniques have been introduced to isolate useful bacteria from the environment, we are still highly dependent on the conventional direct cultivation method to attain pure cultures. However, efficient bacterial isolation is hindered by several factors, including the presence of impurities. In this work, to demonstrate the significance of removing impurities and their impact on bacterial isolation, we employed two approaches: dielectrophoresis (DEP) and fluorescent D-amino acids (FDAA). We successfully attained clean bacterial fractions applicable for downstream processing using these approaches, uniquely designed to identify bacteria based on their characteristics and features. The diversity of bacteria attained by both approaches was investigated using 16S rRNA sequencing and compared to that attained by the standard differential centrifugation method. In addition, the viability of the isolates was also determined via direct cultivation. As a result, the separation of bacteria from impurities allowed for the identification of novel and useful bacteria unique to each approach. Successful cultivation also suggested that both approaches were applicable for attaining viable bacteria. In conclusion, removing impurities to attain clean bacterial fractions promotes the isolation of novel bacteria and thus could aid in the successful isolation of useful bacteria within complex environmental samples.

Optoelectronic Tweezers Micro-Well System for Highly Efficient Single-Cell Trapping, Dynamic Sorting, and Retrieval.

Single-cell arrays have emerged as a versatile method for executing single-cell manipulations across an array of biological applications. In this paper, an innovative microfluidic platform is unveiled that utilizes optoelectronic tweezers (OETs) to array and sort individual cells at a flow rate of 20µLmin-1. This platform is also adept at executing dielectrophoresis (DEP)-based, light-guided single-cell retrievals from designated micro-wells. This presents a compelling non-contact method for the rapid and straightforward sorting of cells that are hard to distinguish. Within this system, cells are individually confined to micro-wells, achieving an impressive high single-cell capture rate exceeding 91.9%. The roles of illuminating patterns, flow velocities, and applied electrical voltages are delved intoin enhancing the single-cell capture rate. By integrating the OET system with the micro-well arrays, the device showcases adaptability and a plethora of functions. It can concurrently trap and segregate specific cells, guided by their dielectric signatures. Experimental results, derived from a mixed sample of HepG2 and L-O2 cells, reveal a sorting accuracy for L-O2 cells surpassing 91%. Fluorescence markers allow for the identification of sequestered, fluorescence-tagged HepG2 cells, which can subsequently be selectively released within the chip. This platform's rapidity in capturing and releasing individual cells augments its potential for future biological research and applications.

On-Chip Micro-Supercapacitor with High Areal Energy Density Based on Dielectrophoretic Assembly of Nanoporous Metal Microwire Electrodes.

Advances in the Internet of Things (IoT) technology have driven the demand for miniaturized electronic devices, prompting research on small-scale energy-storage systems. Micro-supercapacitors (MSCs) stand out in this regard because of their compact size, high power density, high charge-discharge rate, and extended cycle life. However, their limited energy density impedes commercialization. To resolve this issue, a simple and innovative approach is reported herein for fabricating highly efficient on-chip MSCs integrated with nanoporous metal microwires formed by dielectrophoresis (DEP)-driven gold nanoparticle (AuNP) assembly. Placing a water-based AuNP suspension onto interdigitated electrodes and applying an alternating voltage induces in-plane porous microwire formation in the electrode gap. The DEP-induced AuNP assembly and the gold microwire (AuMW) growth rate can be adjusted by controlling the applied alternating voltage and frequency. The microwire-integrated MSC (AuMW-MSC) electrically outperforms its unmodified counterpart and exhibits a 30% larger electrode area, along with 72% and 78% higher specific and areal capacitances, respectively, than a microwire-free MSC. Additionally, AuMW-MSC achieves maximum energy and power densities of 3.33 µWh cm-2 and 2629 µW cm-2, respectively, with a gel electrolyte. These findings can help upgrade MSCs to function as potent energy-storage devices for small electronics.

Quantitative evaluation of dielectrophoretic separation efficiency of cancer exosomes based on fluorescence imaging

Liquid biopsies for cancer diagnosis using exosomes have been studied. Separating cancer cell-derived exosomes from a sample mixture is crucial, especially when they do not dominate the sample at an early stage of cancer. A method based on their electrical properties could provide a simpler separation than immune-affinity methods. We have identified the differences in electrical properties of cancerous exosomes from those of normal ones, suggesting the possibility for their dielectrophoresis (DEP) based separation. This study separated cancerous exosomes from a mixture of normal exosomes. The DEP of fluorescence-labeled exosomes suspended in aqueous media with varying electrical conductivities was investigated. The observed DEP was quantitatively evaluated using the normalization method accounting for the fluorescence fading effect. The results showed that the cancerous exosomes were preferentially concentrated under the appropriate conditions. This suggests that DEP can enable the selective capture of cancer exosomes without chemical labeling.

Optimization of upstream particle concentration from flow using AC electro-osmosis and dielectrophoresis.

There are many applications where upstream sample processing is required to concentrate dispersed particles in flow; this may be to increase the concentration (e.g., to enhance biosensor accuracy) or to decrease it (e.g., by removing contaminants from flow). The AC electrokinetic phenomenon, dielectrophoresis (DEP), has been used widely for particle trapping for flow, but the magnitude of the force drops reduces rapidly with distance from electrode edges, so that nm-scale particles such as viruses and bacteria are only trapped when near the electrode surface. This limits the usable flow rate in the device and can render the final device unusable for practical applications. Conversely, another electrokinetic phenomenon, AC electro-osmosis (ACEO), can be used to move particles to electrode surfaces but is unable to trap them from flow, limiting their ability for sample cleanup or trap-and-purge concentration. In this paper, we describe the optimization of ACEO electrodes aligned parallel to pressure-driven flow as a precursor/preconditioner to capture particles from a flow stream and concentrate them adjacent to the channel wall to enhance DEP capture. This is shown to be effective at flow rates of up to 0.84 ml min-1. Furthermore, the analysis of the 3D flow structure in the ACEO device by both simulation and confocal microscopy suggests that while the system offers significant benefits, the flow structure in the volume near the channel lid is such that while substantial trapping can occur, particles in this part of the chamber cannot be trapped, independent of the chamber height.

Numerical Simulations of Combined Dielectrophoresis and Alternating Current Electrothermal Flow for High-Efficient Separation of (Bio)Microparticles.

High-efficient separation of (bio)microparticles has important applications in chemical analysis, environmental monitoring, drug screening, and disease diagnosis and treatment. As a label-free and high-precision separation scheme, dielectrophoresis (DEP) has become a research hotspot in microparticle separation, especially for biological cells. When processing cells with DEP, relatively high electric conductivities of suspending media are sometimes required to maintain the biological activities of the biosample, which results in high temperature rises within the system caused by Joule heating. The induced temperature gradient generates a localized alternating current electrothermal (ACET) flow disturbance, which seriously impacts the DEP manipulation of cells. Based on this, we propose a novel design of the (bio)microparticle separator by combining DEP with ACET flow to intensify the separation process. A coupling model that incorporates electric, fluid flow, and temperature fields as well as particle tracking is established to predict (bio)microparticle trajectories within the separator. Numerical simulations reveal that both ACET flow and DEP motion act in the same plane but in different directions to achieve high-precision separation between particles. This work provides new design ideas for solving the very tricky Joule heating interference in the DEP separation process, which paves the way for further improving the throughput of the DEP-based (bio)microparticle separation system.

The Effect of Different System Parameters on the Movement of Microbial Cells Using Light-Induced Dielectrophoresis.

The manipulation of single particles remains a topic of interest with many applications. Here we characterize the impact of selected parameters on the motion of single particles thanks to dielectrophoresis (DEP) induced by visible light, in a technique called Light-induced Dielectrophoresis, or LiDEP, also known as optoelectronic tweezers, optically induced DEP, and image-based DEP. Baker's yeast and Candida cells are exposed to an electric field gradient enabled by shining a photoconductive material with a specific pattern of visible light, and their response is measured in terms of the average cell velocity towards the gradient. The impact on cell velocity when varying the shape and color of the light pattern, as well as the distance from the cell to the pattern, is presented. The experimental setup featured a commercial light projector featuring digital light processing (DLP) technology but mechanically modified to accommodate a 40× microscope objective lens. The minimal resolution achieved on the light pattern was 8 µm. Experimental results show the capability for single cell manipulation and the possibility of using different shapes, colors, and distances to determine the average cell velocity.

Bidirectional and Stepwise Rotation of Cells and Particles Using Induced Charge Electroosmosis Vortexes.

The rotation of cells is of significant importance in various applications including bioimaging, biophysical analysis and microsurgery. Current methods usually require complicated fabrication processes. Herein, we proposed an induced charged electroosmosis (ICEO) based on a chip manipulation method for rotating cells. Under an AC electric field, symmetric ICEO flow microvortexes formed above the electrode surface can be used to trap and rotate cells. We have discussed the impact of ICEO and dielectrophoresis (DEP) under the experimental conditions. The capabilities of our method have been tested by investigating the precise rotation of yeast cells and K562 cells in a controllable manner. By adjusting the position of cells, the rotation direction can be changed based on the asymmetric ICEO microvortexes via applying a gate voltage to the gate electrode. Additionally, by applying a pulsed signal instead of a continuous signal, we can also precisely and flexibly rotate cells in a stepwise way. Our ICEO-based rotational manipulation method is an easy to use, biocompatible and low-cost technique, allowing rotation regardless of optical, magnetic or acoustic properties of the sample.

Controllable and Scaffold‐Free Formation of 3D Multicellular Architectures Using a Bipolar Electrode Array

AbstractThe natural environment of cells in vivo is a 3D network in which cellular responses are determined by influences from surrounding cells on all sides. This complex system is hard to be recapitulated in a conventional way. It is highly desirable to replace 2D models with 3D cell culture models. Microfluidic technology offers a variety of advantageous approaches for the long‐term 3D cell culture. Herein, a new cell patterning approach for generating cell clusters of specified size and shape by exploiting induced charge electroosmosis (ICEO) flow at a bipolar electrode array is exploited. Cell or particle clusters can be produced at the edges or centers of bipolar electrodes (BPEs) with controlled organization, by designing electrodes with arbitrary shapes. This platform is further extended to flexibly organize biological units with different properties via the ICEO or dielectrophoresis (DEP) based assembly. Heterotypic cell clusters can be produced at the bipolar electrode (BPE) by adjusting the applied frequencies and the sample injection sequence. This hybrid approach by integrating ICEO flow vortexes and DEP properties of the cells, allows the wireless generation of high‐throughput 3D clusters or spheroids with size adjustability and biocompatibility, which shows great potential in tissue engineering, drug discovery, and tumor research.

Capillarity Enabled Large-Array Liquid Metal Electrodes for Compact and High-Throughput Dielectrophoretic Microfluidics.

Dielectrophoresis (DEP) particle separation has label-free, well-controllable, and low-damage merits. Sidewall microelectrodes made of liquid metal alloy (LMA) inherits the additional advantage of thick electrodes to generate impactful DEP force. However, existing LMA electrode-based devices lack the ability to integrate large-array electrodes in a compact footprint, severely limiting flow rate and thus throughput. Herein, a facile and versatile method is proposed to integrate high-density thick LMA electrodes in microfluidic devices, taking advantage of the passive control ability of capillary burst valves (CBVs). CBVs with carefully designed burst pressures are co-designed in microfluidic channels, allowing self-assembly of LMA electrode array through simple hand-push injection. The arrayed electrode configuration brings the accumulative DEP deflection effect. Specifically, The fabricated 5000 pairs of sidewall electrodes in a compact chip are demonstrted to achieve ten times higher throughput in DEP deflection. The 5000-electrode-pair device is applied to successfully separate four mixed samples, including human peripheral blood mononuclear cells and A549 cellswith the flow rate of 70 µL min-1. It is envisioned that this work can greatly facilitate LMA electrode array fabrication and offer a robust and versatile platform for DEP separation applications.