Negative Dielectrophoretic Force Research Articles

The electrodynamic analysis and manipulation of PEGDA semicircular tubular structures based on optically induced dielectrophoresis

The manipulation of micron-scale semicircular tubular structures has wide applications in micro-nano processing, device manufacturing, biomedicine, and micron sensing and measurement. Here, we propose a method to fabricate and manipulate semicircular tubular structures based on optically induced dielectrophoresis (ODEP). First, electric field intensity simulations are performed for polyethylene glycol diacrylate (PEGDA) semicircular tubular structures with different conductivities and of different heights. In addition, the polarization model based on slender rods reveals that the semicircular tubular structure is subject to a negative dielectrophoretic force and tends to move along the vertical direction of the central axis. Finally, according to the maximum movement speed of the semicircular tubular structure, the resistance and dielectrophoretic force it receives are characterized. This allows for the realization of the translation and rotation operations of semicircular tubular structures of different lengths, and the assembly of multiple structures into different shapes. This assembly method holds significant promise for applications in biomedicine and the manufacturing and processing of micro-nano devices.

Clinical application of serological Alzheimer's disease diagnosis using a highly sensitive biosensor with hydrogel-enhanced dielectrophoretic force

Analysis of a ratio between amyloid beta 1–40 and 1–42 (Aβ1-40 and Aβ1-42) presented in plasm enables a highly accurate diagnosis of Alzheimer's disease (AD). However, the analysis of plasma Aβs is not routinely conducted because of the lack of Aβ detection techniques sensitive enough to specifically detect Aβ from thousands of biomaterials present in the plasma. We developed a hydrogel-patterned spiral microelectrode sensor combined with a hopping dielectrophoretic (DEP) force, combining the negative DEP and positive DEP forces, for Aβ detection. The hydrogel effectively increased the number of immobilized fragmented antibodies in the reaction region of the sensor and enabled size-exclusive passive filtration of non-specific plasma proteins from that region. The hopping DEP force further concentrated the Aβs and removed the non-specific plasma proteins. Consequently, our sensor achieved a limit of detection (LOD) of approximately ∼ 0.15 pg/mL for both Aβ1-40 and Aβ1-42 in the standard plasma. Finally, comparing the ratio between Aβ1-40 and Aβ1-42 signals, we distinguished AD patients from cognitively normal subjects with 95.83% accuracy and 92.31% precision (n = 24, p < 0.0001, One-way ANOVA).

Cell Washing and Solution Exchange in Droplet Microfluidic Systems.

Water-in-oil emulsion droplet microfluidic systems have been extensively developed, and currently, almost all cell handling steps can be conducted in this format. An exception is the cell washing and solution exchange step, which is commonly utilized in many conventional cell assays. This paper presents an in-droplet cell washing and solution exchange technology that utilizes dielectrophoretic (DEP) force to move all cells to one side of a droplet, followed by asymmetrical splitting of the droplet to obtain a small daughter droplet that contains all or most of the cells, and then finally merges this cell-concentrated droplet with a new droplet that contains the desired solution. These sequential droplet manipulation steps were integrated into a single platform, where up to 88% of the original solution in the droplet could be exchanged with the new solution while keeping cell loss to less than 5%. Two application examples were demonstrated using the developed technology. In the first example, green microalga Chlamydomonas reinhardtii cells were manipulated using negative DEP force to exchange the regular culture medium with a nitrogen-limited medium to induce lipid production. In the second example, Salmonella enterica cells were manipulated using positive DEP force to replace fluorescent dye that models fluorescent cell stains that contribute to high background noise in fluorescence-based droplet content detection with fresh buffer solution, significantly improving the droplet content detection sensitivity. Since the cell washing step is one of the most frequently utilized steps in many cell biology assays, we expect that the developed technology can significantly broaden the type of assay that can be conducted in droplet microfluidic format.

Bacteria and cancer cell pearl chain under dielectrophoresis.

In this work, we aim to observe and study the physics of bacteria and cancer cells pearl chain formation under dielectrophoresis (DEP). Experimentally, we visualized the formation of Bacillus subtilis bacterial pearl chain and human breast cancer cell (MCF-7) chain under positive and negative dielectrophoretic force, respectively. Through a simple simulation with creeping flow, AC/DC electric fields, and particle tracing modules in COMSOL, we examined the mechanism by which bacteria self-organize into a pearl chain across the gap between two electrodes via DEP. Our simulation results reveal that the region of greatest positive DEP force shifts from the electrode edge to the leading edge of the pearl chain, thus guiding the trajectories of free-flowing particles toward the leading edge via positive DEP. Our findings additionally highlight the mechanism why the free-flowing particles are more likely to join the existing pearl chain rather than starting a new pearl chain. This phenomenon is primarily due to the increase in magnitude of electric field gradient, and hence DEP force exerted, with the shortening gap between the pearl chain leading edge and the adjacent electrode. The findings shed light on the observed behavior of preferential pearl chain formation across electrode gaps.

Computational Modeling of Dielectrophoretic Microfluidic Channel for Simultaneous Separation of Red Blood Cells and Platelets

Background: In this paper, the design and computational modeling of microfluidic channel capable of separating platelets and Red Blood Cell (RBC) from the other blood cells are proposed. Materials and Methods: Separation based on their sizes is made possible by utilizing negative dielectrophoretic (n-DEP) force in fusion with drag force. An array of 38º angled electrode separated by 70 μm distance is designed within the microchannel and analyzed for non-uniform electric field distribution. Results and Conclusion: The molecule movement within the microchannel under induced electric field is simulated to demonstrate the separation using the particle trajectories module. A numerical study is performed for the calculation of Clausius Mossotti (CM) factor, n-DEP force and drag force.

Dielectrophoretic separation of monocytes from cancer cells in a microfluidic chip using electrode pitch optimization.

This study proposes a microfluidic device capable of separating monocytes from a type of cancer cell that is called T-cell acute lymphoblastic leukemia (RPMI-8402) in a continuous flow using negative and positive dielectrophoretic forces. The use of both the hydrodynamic and dielectrophoretic forces allows the separation of RPMI-8402 from monocytes based on differences in their intrinsic electrical properties and sizes. The specific crossover frequencies of monocytes and RPMI-8402 cells have been obtained experimentally. The optimum ranges of electrode pitch-to-channel height ratio at the cross sections with different electrode widths have been generally calculated by numerical simulations of the gradients of the electric field intensities and calculation their effective values (root-mean-square). In the device, the cell sorting has been conducted empirically, and then, the separation performance has been evaluated by analyzing the images before and after dielectrophoretic forces applied to the cells. In this work, the design of a chip with 77μm gold-titanium electrode pitch was investigated to achieve high purity of monocytes of 95.2%. The proposed device can be used with relatively low applied voltages, as low as 16.5V (peak to peak). Thus, the design can be used in biomedical diagnosis and chemical analysis applications as a lab-on-chip platform. Also, it can be used for the separation of biological cells such as bacteria, RNA, DNA, and blood cells.

An Improved Escherichia Coli Bacterium Detection in Microchannel Based on Dielectrophoresis Impedance Measurements

In this study, a bacterial detection technique utilizing benefits of both positive and negative dielectrophoresis has been simulated with COMSOL Multiphysics. The two dimensional proposed model has six microelectrodes across microfluidic channel. Four of them were positioned on the upper side and the rest were positioned on the bottom side of the microchannel, which serve as a bacteria concentrator using negative dielectrophoresis and as a bacteria detector using positive dielectrophoresis, respectively. In this simulation, the target particle is Escherichia coli that was flowing into the microchannel and repelled under negative dielectrophoretic force exerted by the upper side microelectrodes, and were pushed toward the bottom side microelectrodes situated at the downstream. Finally, concentrated bacteria have been captured and detected by dielectrophoretic impedance measurement method. The numerical simulation proved that negative dielectrophoretic force could increase sensitivity with respect to the absence of the negative dielectrophoretic force and eventually, results represented that 80 percent of releasing E. coli bacteria trapped near bottom side microelectrodes.

Measurement of the Imaginary Part of the Clausius-Mossotti Factor of Particle/Cell via Dual Frequency Electrorotation.

A simple and inexpensive method using planar electrodes was proposed for the measurement of the imaginary part of the Clausius-Mossotti factor, , of particle/cell for electrorotation (ER) and travelling wave dielectrophoresis (twDEP). It is based on the balance between the dielectrophoretic and viscous torques on a particle undergoing ER subject to dual frequency operation in an ER chamber. A four-phase ac voltage signal with a given frequency is applied for generating ER for measurement, and another two-phase signal is applied at a selected frequency for generating a negative dielectrophoretic force for confining the particle motion, instead of using laser tweezer or three-dimensional electrodes in the literature. Both frequencies can be applied to the same electrodes in a four-electrode ER system and to alternative different electrodes in an eight-electrode ER system, and both systems are capable for providing accurate measurement. The measurements were validated by comparing with the theoretical result using sephadex particles in KCl solution, and with the existing experimental results for various human cancer cells in medium with conductivity from 0.01–1.2 S/m, using ER with optical tweezer and dual frequency twDEP. Contrast between the ER and the twDEP methods (the current two available methods) was discussed and commented. The present method could provide measurement for wider frequency range and more accurate result near = 0, in comparison with the results using the twDEP method. However, the twDEP method could perform much more rapid measurement. Detailed forces and torque were calculated inside the ER chamber for understanding the physics and assessing the characteristics of the dual frequency ER method. This study is of academic interest as the torque in ER and the force in twDEP can be calculated only when is known. It also finds biomedical applications as the -spectra can be served as physical phenotypes for different cells, and can be applied for deriving dielectric properties of cells.

Selective Retrieval of Individual Cells from Microfluidic Arrays Combining Dielectrophoretic Force and Directed Hydrodynamic Flow.

Hydrodynamic-based microfluidic platforms enable single-cell arraying and analysis over time. Despite the advantages of established microfluidic systems, long-term analysis and proliferation of cells selected in such devices require off-chip recovery of cells as well as an investigation of on-chip analysis on cell phenotype, requirements still largely unmet. Here, we introduce a device for single-cell isolation, selective retrieval and off-chip recovery. To this end, singularly addressable three-dimensional electrodes are embedded within a microfluidic channel, allowing the selective release of single cells from their trapping site through application of a negative dielectrophoretic (DEP) force. Selective capture and release are carried out in standard culture medium and cells can be subsequently mitigated towards a recovery well using micro-engineered hybrid SU-8/PDMS pneumatic valves. Importantly, transcriptional analysis of recovered cells revealed only marginal alteration of their molecular profile upon DEP application, underscored by minor transcriptional changes induced upon injection into the microfluidic device. Therefore, the established microfluidic system combining targeted DEP manipulation with downstream hydrodynamic coordination of single cells provides a powerful means to handle and manipulate individual cells within one device.

Continuous Cell Characterization and Separation by Microfluidic Alternating Current Dielectrophoresis.

A novel alternating current (ac)-dielectrophoretic (DEP) microfluidic chip for continuous cell characterization and separation is presented in this paper. To generate DEP forces, two electrode-pads are embedded in a set of asymmetric orifices on the opposite sidewalls to produce the nonuniform electric fields. In the vicinity of a small orifice, the cells experience the strongest nonuniform gradient and are drawn toward it by the positive DEP forces, while the cells experiencing a negative DEP force are repelled away and move toward the large orifice. The DEP behaviors of yeast cells in suspending media with different ionic concentrations, i.e., different electrical conductivities, and over a large range of the ac electric field frequency were investigated. Furthermore, the lateral migrations of yeast cells as a function of the ac frequency were measured. The trends of measured lateral migrations of yeast cells are similar to the corresponding Clausius-Mossotti (CM) factors. In addition, by adjusting the frequency and strength of the ac electric field, the continuous separation of live and dead yeast cells as well as the yeast cells with targeted diameter and dielectric property can be easily achieved. This is the first time that the measurement of ac-DEP lateral migration of yeast cells in solutions with different electrical conductivities as a function of the applied frequency in a microfluidic chip was reported. This ac-DEP system provides a method to characterize the crossover frequency of the specific cells and manipulate the targeted cells.

Multiplexing microelectrodes for dielectrophoretic manipulation and electrical impedance measurement of single particles and cells in a microfluidic device.

This work presents a microfluidic device, which was patterned with (i) microstructures for hydrodynamic capture of single particles and cells, and (ii) multiplexing microelectrodes for selective release via negative dielectrophoretic (nDEP) forces and electrical impedance measurements of immobilized samples. Computational fluid dynamics (CFD) simulations were performed to investigate the fluidic profiles within the microchannels during the hydrodynamic capture of particles and evaluate the performance of single-cell immobilization. Results showed uniform distributions of velocities and pressure differences across all eight trapping sites. The hydrodynamic net force and the nDEP force acting on a 6μm sphere were calculated in a 3D model. Polystyrene beads with difference diameters (6, 8, and 10μm) and budding yeast cells were employed to verify multiple functions of the microfluidic device, including reliable capture and selective nDEP-release of particles or cells and sensitive electrical impedance measurements of immobilized samples. The size of immobilized beads and the number of captured yeast cells can be discriminated by analyzing impedance signals at 1MHz. Results also demonstrated that yeast cells can be immobilized at single-cell resolution by combining the hydrodynamic capture with impedance measurements and nDEP-release of unwanted samples. Therefore, the microfluidic device integrated with multiplexing microelectrodes potentially offers a versatile, reliable, and precise platform for single-cell analysis.

Multi-Stage Particle Separation based on Microstructure Filtration and Dielectrophoresis.

Particle separation is important in chemical and biomedical analysis. Among all particle separation approaches, microstructure filtration which based particles size difference has turned into one of the most commonly methods. By controlling the movement of particles, dielectrophoresis has also been widely adopted in particle separation. This work presents a microfluidic device which combines the advantages of microfilters and dielectrophoresis to separate micro-particles and cells. A three-dimensional (3D) model was developed to calculate the distributions of the electric field gradient at the two filter stages. Polystyrene particles with three different sizes were separated by micropillar array structure by applying a 35-Vpp AC voltage at 10 KHz. The blocked particles were pushed off the filters under the negative dielectrophoretic force and drag force. A mixture of Haematococcus pluvialis cells and Bracteacoccus engadinensis cells with different sizes were also successfully separated by this device, which proved that the device can separate both biological samples and polystyrene particles.

Dielectrophoretic choking phenomenon in a converging-diverging microchannel for Janus particles.

The dielectrophoretic (DEP) choking phenomenon is revisited for Janus particles that are transported electrokinetically through a microchannel constriction by a direct-current (DC) electric field. The negative DEP force that would block a particle with a diameter significantly smaller than that of the constriction at its inlet is seen to be relaxed by the rotation of the Janus particle in a direction that minimizes the magnitude of the DEP force. This allows the particle to pass through the constriction completely. An arbitrary Lagrangian-Eulerian (ALE) numerical method is used to solve the nonlinearly coupled electric field, flow field, and moving particle, and the DEP force is calculated by the Maxwell stress tensor (MST) method. The results show how Janus particles with non-uniform surface potentials overcome the DEP force and present new conditions for the DEP choking by a parametric study. Particle transportation through microchannel constrictions is ubiquitous, and particle surface properties are more likely to be non-uniform than not in practical applications. This study provides new insights of importance for non-uniform particles transported electrokinetically in a microdevice.

Microfluidic platform for dielectrophoretic separation of bio-particles using serpentine microelectrodes

Recently, there is a great concern in developing integrated microfluidic platforms for biological analysis systems that can perform general biochemistry and biological analysis. Dielectrophoresis, the movement of particles in a non-uniform electric field is widely used to separate viable and non viable cells with good efficiency. The present study aims to develop a universal bio-particle separator in a C-serpentine geometry which could separate particles by varying the operating conditions. We present the design, fabrication and testing for continuous separation structure based on dielectrophoretic forces. The C-Serpentine geometry is composed of consecutive C shaped curves to generate a gradient required for the dielectrophoretic effect. Known mixture of viable and non viable cells of Saccharomyces Cerevisiae was selectively trapped using negative dielectrophoretic force generated by microelectrodes. Through measurement of cell count, a trapping efficiency of 86% was obtained for mixed cell suspension and 93–98% trapping efficiency was obtained in case of trapping live and dead yeast cells individually. Also the results show that cell viability was not affected by the separation procedure as percentage of viable cells was calculated to be 96%.

Lateral fluid flow fractionation using dielectrophoresis (LFFF-DEP) for size-independent, label-free isolation of circulating tumor cells

This short communication introduces a continuous-flow, dielectrophoresis-based lateral fluid flow fractionation microdevice for detection/isolation of circulating tumor cells in the presence of other haematological cells. The device utilizes two sets of planar interdigitated transducer electrodes micropatterned on top of a glass wafer using standard microfabrication techniques. A microchannel with a single inlet and two outlets, realized in polydimethylsiloxane, is bonded on the glass substrate. The two sets of electrodes slightly protrude into the microchannel. Both of the electrode sets are energized with signals at different frequencies and different operating voltages ensuring that the cancer cells experience positive dielectrophoretic force from one set of the electrodes and negative dielectrophoretic force from the other array. Normal cells experience unequal negative dielectrophoretic forces from opposing sets of electrodes. The resultant dielectrophoretic forces on cancer and normal cells push them to flow towards their designed outlets. Successful isolation of green fluorescent protein-labelled MDA-MB-231 breast cancer cells from regular blood cells, both suspended in a sucrose/dextrose medium, is reported in this work.

Selective position of individual cells without lysis on a circular window array using dielectrophoresis in a microfluidic device

We trapped individual cells between two circular windows using negative dielectrophoretic (DEP) force and then sequentially trapped them inside circular windows by positive DEP force without electrical lysis in a microfluidic device. Three parameters, (1) the transmembrane potential that determines the lysis of a cell, (2) individual cell size that affects the trapped position accuracy of the cell, and (3) the Clausius–Mossotti (CM) factor that decides the trapped efficiency of the cell, were characterized experimentally and numerically in this sequential cell trapping technique. In this characterization, we confirmed that the swap rate of applied voltage frequency, size similarity between the cell and circular window, and instantaneous change rate of Re(f CM) as a function of frequency were important factors in determining the selective position of individual cells without lysis. Our results provide useful suggestions for designing the structure of microfluidic DEP devices and optimizing variables required to manipulate individual cell trapping using both negative and positive DEP forces.

DC biased low-frequency insulating constriction dielectrophoresis for protein biomolecules concentration

Sample enrichment or molecules concentration is considered an essential step in sample processing of miniaturized devices aimed at biosensing and bioanalysis. Among all the means involved to achieve this aim, dielectrophoresis (DEP) is increasingly employed in molecules manipulation and concentration because it is non-destructive and high efficiency. This paper presents a methodology to achieve protein concentration utilizing the combination effects of electrokinetics and low frequency insulating dielectrophoresis (iDEP) generated within a microfluidic device, in which a submicron constricted channel was fabricated using DNA molecular combing and replica molding. This fabrication technique avoids using e-beam lithography or other complicated nanochannel fabrication methods, and provides an easy and low cost approach with the flexibility of controlling channel dimensions to create highly constricted channels embedded in a microfluidic device. With theoretical analysis and experiments, we demonstrated that fluorescein isothiocyanate conjugated bovine serum albumin (FITC-BSA) protein molecules can be significantly concentrated to form an arc-shaped band near the constricted channel under the effects of a negative dielectrophoretic force and DC electrokinetic forces within a short period of time. It was also observed that the amplitudes of the applied DC and AC electric fields, the AC frequencies as well as the suspending medium conductivities had strong effects on the concentration responses of the FITC-BSA molecules, including the concentrated area and position, intensities of the focused molecules, and concentration speed. Our method provides a simple and flexible approach for quickly concentrating protein molecules by controlling the applied electric field parameters. The iDEP device reported in this paper can be used as a stand-alone sensor or worked as a pre-concentration module integrated with biosensors for protein biomarker detection. Furthermore, low frequency dielectrophoresis provides practical uses for integrating the concentration module with a portable biosensing system.

CMOS single cell dielectrophoresis cytometer

An integrated dielectrophoresis cytometer has been designed and implemented using a standard 0.35μm CMOS technology. This design makes use of differential ring oscillators. The frequency difference between the two oscillators, in presence of a cell, is detected by an XOR gate. The frequency difference is then measured by a frequency counter. This sensor has ∼14aF capacitance sensitivity at ∼1.4GHz. It is used for detection of 10–20μm polystyrene spheres and Chinese Hamster Ovary cells using coplanar microelectrodes on top of the CMOS chip. The design demonstrates that it is possible to differentiate between particles experiencing an altitude change resulting from either a positive or negative dielectrophoretic force in a microfluidic channel. The total area occupied by the DEP system is 0.6mm2. This sensor can be used in high throughput applications with a density of more than 160DEP sensor units in 1cm2. The sensor integrates all the required parts of a DEP cytometer -excluding the pumping system and frequency counter-, making a semi-integrated lab on chip system for single cell detection and analysis.

Microparticle trajectories in a high-throughput channel for contact-free fractionation by dielectrophoresis

Continuous, contact-free fractionation of sensitive microparticles at high throughput is a challenge. For this purpose, we developed a sheath flow assisted dielectrophoretic (DEP) field-flow separator with a tailored arrangement of cylindrical interdigitated electrodes (cIDE) and observed size-dependent trajectories of dispersed particles. Using a voltage input of 200Veff at a frequency of 200kHz, polystyrene particles (45, 25, and 11µm in diameter) levitated to different heights along the channel length due to a negative DEP force. Experimental observations agree well with simulated particle trajectories that were obtained by a modified Lagrangian particle tracking model in combination with Laplace's and Navier–Stokes equations. By exploiting the size-dependent levitation height difference the desired particle size fraction can be collected at a specific channel length. The required channel length of the proposed cIDE separator increases with decreasing particle size to be separated. The quality of theoretical fractionation, which we quantify by resolution, improves strongly with reduced collector width, reduced volume flow rate and increased voltage input. The sensitivity of these dependencies increases with decreasing particle size. We calculated a theoretical throughput of up to 47mLmin−1 when trading-off design and operation parameters, allowing for contact-free fractionation of sensitive microparticles with negligible shear stress.

Sensitivity improvement of an electrical sensor achieved by control of biomolecules based on the negative dielectrophoretic force

Effective control of nano-scale biomolecules can enhance the sensitivity and limit of detection of an interdigitated microelectrode (IME) sensor. Manipulation of the biomolecules by dielectrophoresis (DEP), especially the negative DEP (nDEP) force, so that they are trapped between electrodes (sensing regions) was predicted to increase the binding efficiency of the antibody and target molecules, leading to a more effective reaction. To prove this concept, amyloid beta 42 (Aβ42) and prostate specific antigen (PSA) protein were respectively trapped between the sensing region owing to the nDEP force under 5V and 0.05V, which was verified with COMSOL simulation. Using the simulation value, the resistance change (ΔR/Rb) of the IME sensor from the specific antibody–antigen reaction of the two biomolecules and the change in fluorescence intensity were compared in the reference (pDEP) and nDEP conditions. The ΔR/Rb value improved by about 2-fold and 1.66-fold with nDEP compared to the reference condition with various protein concentrations, and these increases were confirmed with fluorescence imaging. Overall, nDEP enhanced the detection sensitivity for Aβ42 and PSA by 128% and 258%, respectively, and the limit of detection improved by up to 2-orders of magnitude. These results prove that DEP can improve the biosensor's performance.