Sheath-assisted versus sheathless dielectrophoretic particle separation

Lab-on-chip devices are widely being used for binary and ternary cell/particle separation applications. Among the lab-on-chip methods, dielectrophoresis (DEP) is a cost-effective and label-free method, with great capabilities for size-based separation of cells and particles, which is mostly performed in sheath-assisted forms. However, the elimination of the sheath flows offers advantages such as ease of operation and higher sample throughput. In this work, we present a comparison of sheath-assisted and sheathless DEP separation of three sizes of microparticles using tilted electrodes. The sheath-assisted design was capable of separating the 5, 10, and 15 μm particles with a separation efficiency as high as 98.0% for 15 μm particles. By adding a DEP focusing region, a sheathless DEP separator was proposed, which offered higher throughputs (up to 10 times) at the cost of lowering the separation efficiency (a reduction up to 10.3% for 15 μm) compared to the sheath-assisted design. To enhance the separation efficiency, a combination of the DEP focusing accompanied by weak sheath flows from both sides was proposed. This design achieved the highest sample separation yield in the outlets (as high as 98.7% for 15 μm) with a sample throughput of more than 4.2 μL/min. This study provides insights into the choice of an appropriate platform for any application in which the yield, purity, throughput, and portability must be considered.     

Size and medium conductivity dependence on dielectrophoretic behaviors of gas core poly‐l‐lysine shell nanoparticles

Dynamic (dis)assembly of biocompatible nanoparticles into 3D, packed structures would benefit drug delivery, films, and diagnostics. Dielectrophoretic (DEP) microdevices can rapidly assemble and manipulate polarizable particles within nonuniform electric fields. DEP has primarily discerned micrometer particles since nanoparticles experience smaller forces. This work examines conductivity and size DEP dependencies of previously unexplored spherical core‐shell nanoparticle (CSnp) into 3D particle assemblies. Poly‐l ‐lysine shell material was custom synthesized around a gas core to form CSnps. DEP frequencies from 1 kHz to 80 MHz at fixed 5 volts peak‐to‐peak and medium conductivities of 10^?5 and 10^?3 S/m were tested. DEP responses of ～220 and ～400 nm poly‐l ‐lysine CSnps were quantified via video intensity densitometry at the microdevice's quadrapole electrode center for negative DEP (nDEP) and adjacent to electrodes for positive DEP. Intensity densitometry was then translated into a relative DEP response curve. An unusual nDEP peak occurred at ～57 MHz with 25–80 times greater apparent nDEP force. All electrical circuit components were then impedance matched, which changed the observed response to weak positive DEP at low frequencies and consistently weak nDEP from ～100 kHz to 80 MHz. This impedance‐matched behavior agrees with conventional Clausius–Mossotti DEP signatures taking into account the gas core's contributions to the polarization mechanisms. This work describes a potential pitfall when conducting DEP at higher frequencies in microdevices and concurrently demonstrates nDEP behavior for a chemically and structurally distinct particle system. This work provides insight into organic shell material properties in nanostructures and strategies to facilitate dynamic nanoparticle assemblies.     

Revisit of wall‐induced lateral migration in particle electrophoresis through a straight rectangular microchannel: Effects of particle zeta potential

Previous studies have reported a lateral migration in particle electrophoresis through a straight rectangular microchannel. This phenomenon arises from the inherent wall‐induced electrical lift that can be exploited to focus and separate particles for microfluidic applications. Such a dielectrophoretic‐like force has been recently found to vary with the buffer concentration. We demonstrate in this work that the particle zeta potential also has a significant effect on the wall‐induced electrical lift. We perform an experimental study of the lateral migration of equal‐sized polystyrene particles with varying surface charges under identical electrokinetic flow conditions. Surprisingly, an enhanced focusing is observed for particles with a faster electrokinetic motion, which indicates a substantially larger electrical lift for particles with a smaller zeta potential. We speculate this phenomenon may be correlated with the particle surface conduction that is a strong function of particle and fluid properties.     

Mapping alternating current electroosmotic flow at the dielectrophoresis crossover frequency of a colloidal probe

AC electroosmotic (ACEO) flow above the gap between coplanar electrodes is mapped by the measurement of Stokes forces on an optically trapped polystyrene colloidal particle. E²‐dependent forces on the probe particle are selected by amplitude modulation (AM) of the ACEO electric field (E) and lock‐in detection at twice the AM frequency. E²‐dependent DEP of the probe is eliminated by driving the ACEO at the probe's DEP crossover frequency. The location‐independent DEP crossover frequency is determined, in a separate experiment, as the limiting frequency of zero horizontal force as the probe is moved toward the midpoint between the electrodes. The ACEO velocity field, uncoupled from probe DEP effects, was mapped in the region 1–9 μm above a 28 μm gap between the electrodes. By use of variously sized probes, each at its DEP crossover frequency, the frequency dependence of the ACEO flow was determined at a point 3 μm above the electrode gap and 4 μm from an electrode tip. At this location the ACEO flow was maximal at ～117 kHz for a low salt solution. This optical trapping method, by eliminating DEP forces on the probe, provides unambiguous mapping of the ACEO velocity field.     

Dielectrophoresis switching with vertical sidewall electrodes for microfluidic flow cytometry

A novel dielectrophoresis switching with vertical electrodes in the sidewall of microchannels for multiplexed switching of objects has been designed, fabricated and tested. With appropriate electrode design, lateral DEP force can be generated so that one can dynamically position particulates along the width of the channel. A set of interdigitated electrodes in the sidewall of the microchannels is used for the generation of non-uniform electrical fields to generate negative DEP forces that repel beads/cells from the sidewalls. A countering DEP force is generated from another set of electrodes patterned on the opposing sidewall. These lateral negative DEP forces can be adjusted by the voltage and frequency applied. By manipulating the coupled DEP forces, the particles flowing through the microchannel can be positioned at different equilibrium points along the width direction and continue to flow into different outlet channels. Experimental results for switching biological cells and polystyrene microbeads to multiple outlets (up to 5) have been achieved. This novel particle switching technique can be integrated with other particle detection components to enable microfluidic flow cytometry systems.     

Continuous dielectrophoretic separation of particles in a spiral microchannel

Particle separation is a fundamental operation in the areas of biology and physical chemistry. A variety of force fields have been used to separate particles in microfluidic devices, among which electric field may be the most popular one due to its general applicability and adaptability. So far, however, electrophoresis‐based separations have been limited primarily to batchwise processes. Dielectrophoresis (DEP)‐based separations require in‐channel micro‐electrodes or micro‐insulators to produce electric field gradients. This article introduces a novel particle separation technique in DC electrokinetic flow through a planar double‐spiral microchannel. The continuous separation arises from the cross‐stream dielectrophoretic motion of particles induced by the non‐uniform electric field inherent to curved channels. Specifically, particles are focused by DEP to one sidewall of the first spiral, and then dielectrophoretically deflected toward the other sidewall of the second spiral at a particle‐dependent rate, leading to focused particle streams along different flow paths. This DEP‐based particle separation technique is demonstrated in an asymmetric double‐spiral microchannel by continuously separating a mixture of 5/10 μm particles and 3/5 μm particles.     

Microfluidic system for dielectrophoretic separation based on a trapezoidal electrode array

This paper presents a novel microfluidic device for dielectrophoretic separation based on a trapezoidal electrode array (TEA). In this method, particles with different dielectric properties are separated by the device composed of the TEA for the dielectrophoretic deflection of particles under negative dielectrophoresis (DEP) and poly(dimethylsiloxane)(PDMS) microfluidic channel with a sinuous and expanded region. Polystyrene microparticles are exposed to an electric field generated from the TEA in the microfluidic channel and are dielectrophoretically focused to make all of them line up to one sidewall. When these particles arrive at the region of another TEA for dielectrophoretic separation, they are separated having different positions along the perpendicular direction to the fluid flow due to their different dielectrophoretic velocities. To evaluate the separation process and performance, both the effect of the flow rate on dielectrophoretic focusing and the influence of the number of trapezoidal electrodes on dielectrophoretic separation are investigated. Now that this method utilizes the TEA as a source of negative DEP, non-specific particle adhering to the electrode surface can be prevented; conventional separation approaches depending on the positive DEP force suffer from this problem. In addition, since various particle types are continuously separated, this method can be easily applicable to the separation and analysis of various dielectric particles with high particle recovery and selectivity.     

Rapid microparticle patterning by enhanced dielectrophoresis effect on a double-layer electrode substrate

We present a feasible dielectrophoresis (DEP) approach for rapid patterning of microparticles on a reusable double-layer electrode substrate in microfluidics. Simulation analysis demonstrated that the DEP force was dramatically enhanced by the induced electric field on top interdigitated electrodes. By adjusting electric field intensity through the bottom electrodes on thin glass substrate (100 μm), polystyrene particles (10 μm) were effectively patterned by top electrodes within several seconds (<5 s). The particle average velocity can reach a maximum value of about 20.0±3.0 μm/s at 1 MHz with the strongest DEP force of 1.68 pN. This approach implements integration of functional electrodes into one substrate and avoids direct electrical connection to biological objects, providing a potential lab-on-chip system for biological applications.     

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.     

Combined detection of AC‐electrokinetic effects: Experiments with three‐axial chicken red blood cells

Dielectrophoresis (DEP), electrorotation (ROT), and electro‐orientation were used for the dielectric spectroscopy of nucleated three‐axial chicken red blood cells (CRBCs). Because the different AC‐electrokinetic effects are not mutually independent, their DEP and ROT spectra were combined in ranges separated by the reorientation of the CRBCs in the inhomogeneous linear DEP and circular ROT fields. This behavior can be qualitatively described by a single‐shell ellipsoidal model. Whereas in linear fields, the maximum of the Clausius–Mossotti factor along the three axes determines the orientated axis, in circular fields, the minimum of the factor determines the axis perpendicularly orientated to the field plane. Quantitatively, it has not been possible to find a consistent parameter set for fitting the DEP and ROT spectra, as well as the reorientation frequencies. Our ellipsoidal CRBC standard model had semiaxes of a = 7.7 μm, b = 4.0 μm, and c = 1.85 μm, a relative permittivity of 35 to 45 and conductivity of 0.36 to 0.04 S/m for the cytoplasm, combined with a specific capacitance of 10 to 14 mF/m² and a conductivity of 3500 S/m² for the cell membrane. The fits in different external conductivity ranges between external conductivities of 0.015 and 1.0 S/m were improved when the membrane capacitance was changed between 4 to 25 mF/m² depending on the method used. A similar transition was reflected in the effective properties of a three‐shell spherical model containing an internal membranous sphere with the geometry of the CRBC nucleus. Our findings suggest that the simultaneous interpretation of various AC‐electrokinetic spectra is a step toward the dielectric fingerprinting of biological cells.     

Breast cancer cell obatoclax response characterization using passivated‐electrode insulator‐based dielectrophoresis

Inherent electrical properties of cells can be beneficial to characterize different cell lines and their response to experimental drugs. This paper presents a novel method to characterize the response of breast cancer cells to drug stimuli through use of off‐chip passivated‐electrode insulator‐based dielectrophoresis (OπDEP) and the application of AC electric fields. This work is the first to demonstrate the ability of OπDEP to differentiate between two closely related breast cancer cell lines, LCC1 and LCC9 while assessing their drug sensitivity to an experimental anti‐cancer agent, Obatoclax. Although both cell lines are derivatives of estrogen‐responsive MCF‐7 breast cancer cells, growth of LCC1 is estrogen independent and anti‐estrogen responsive, while LCC9 is both estrogen‐independent and anti‐estrogen resistant. Under the same operating conditions, LCC1 and LCC9 had different DEP profiles. LCC1 cells had a trapping onset (crossover) frequency of 700 kHz and trapping efficiencies between 30–40%, while LCC9 cells had a lower crossover frequency (100 kHz) and showed higher trapping efficiencies of 40–60%. When exposed to the Obatoclax, both cell lines exhibited dose‐dependent shifts in DEP crossover frequency and trapping efficiency. Here, DEP results supplemented with cell morphology and proliferation assays help us to understand the response of these breast cancer cells to Obatoclax.     

Multistep manipulations of poly(methyl‐methacrylate) submicron particles using dielectrophoresis

A microfluidic chip for multistep manipulations of PMMA submicron particles (PMMA‐SMPs) based on dielectrophoresis (DEP) has been developed that includes four main functions of focusing, guiding, trapping, and releasing the SMPs. The structure of the DEP chip consists of a top electrode made of indium tin oxide, a flow chamber formed by optically clear adhesive tape and bottom electrodes with different patterns for different purposes. The bottom electrodes can be divided into three parts: a fish‐bone‐type electrode array that provides the positive DEP force for focusing the suspended nanoparticles (NPs) near the inlet in the flow chamber; the second is for switching and guiding the focused NPs along the electrode surface to the target area, like a flow passing along a virtual channel; and a trapping electrode in the downstream for trapping and releasing the guided NPs. According to the simulation and experimental results, NPs can be aligned along the electrode of the focusing electrode and guided toward the target electrode by means of a positive DEP force between the top and bottom electrodes, with the effects of Brownian motion and Stokes force. In order to demonstrate the sequence of DEP manipulations, a PMMA‐NP suspension is introduced to the DEP chip; the size of the PMMA‐SMPs is about 300 nm. Furthermore, a LabVIEW program developed for sequence control of the AC signals for the multistep manipulations. Consequently, the DEP chip provides an excellent platform technology for the multistep manipulation of SMPs.     

Effects of dielectrophoresis on thrombogenesis in human whole blood

Thrombogenesis (blood clot formation) is a major barrier to the development of biomedical devices that interface with blood. Although state‐of‐the‐art chemically and pharmacologically mediated clot mitigation strategies are effective, some limitations of such approaches include depletion of active agents, or adverse reactions in patients. Increased clotting protein adsorption and platelet adhesion, which occur when artificial surfaces are exposed to blood result in enhanced clot formation on artificial surfaces. It is hypothesized that repelling proteins and platelets using dielectrophoresis (DEP), a contact‐free particle manipulation technique, will reduce clot formation in biomedical devices. In this paper, the effect of DEP on thrombogenesis in human blood is investigated. Undiluted whole blood from human donors is pumped through microchannels at a physiological shear rate (400 s ⁻¹). Experiments are performed by applying 0 V, 0.5 V_rms , 2 V_rms , and 3 V_rms to electrodes in the channel. Clot formation is observed to decrease in experiments in which DEP electrodes are active (average of 6% coverage @ 0V reduced to 0.08% coverage @ 3 V_rms ). Repulsion is more effective at higher voltages. DEP causes a quantifiable reduction in microscopic and macroscopic clot formation in PDMS microchannels.     

Investigation of viscoelastic focusing of particles and cells in a zigzag microchannel

Microfluidic particle focusing has been a vital prerequisite step in sample preparation for downstream particle separation, counting, detection, or analysis, and has attracted broad applications in biomedical and chemical areas. Besides all the active and passive focusing methods in Newtonian fluids, particle focusing in viscoelastic fluids has been attracting increasing interest because of its advantages induced by intrinsic fluid property. However, to achieve a well-defined focusing position, there is a need to extend channel lengths when focusing micrometer-sized or sub-microsized particles, which would result in the size increase of the microfluidic devices. This work investigated the sheathless viscoelastic focusing of particles and cells in a zigzag microfluidic channel. Benefit from the zigzag structure of the channel, the channel length and the footprint of the device can be reduced without sacrificing the focusing performance. In this work, the viscoelastic focusing, including the focusing of 10 μm polystyrene particles, 5 μm polystyrene particles, 5 μm magnetic particles, white blood cells (WBCs), red blood cells (RBCs), and cancer cells, were all demonstrated. Moreover, magnetophoretic separation of magnetic and nonmagnetic particles after viscoelastic pre-focusing was shown. This focusing technique has the potential to be used in a range of biomedical applications.     

Electrodeless direct current dielectrophoresis using reconfigurable field-shaping oil barriers

We demonstrate dielectrophoretic (DEP) potential wells using pairs of insulating oil menisci to shape the DC electric field. These oil menisci are arranged in a configuration similar to the quadrupolar electrodes, typically used in DEP, and are shown to produce similar field gradients. While the one-pair well produces a focusing effect on particles in flow, the two-pair well results in creating spatial traps against crossflows. Uncharged polystyrene particles were used to map the DEP force fields and the experimental observations were compared against the field profiles obtained by numerically solving Maxwell's equations. We demonstrate trapping of a single particle due to negative DEP against a pressure-driven crossflow. This can be easily extended to trap and hold cells and other objects against flow for a longer time. We also show the results of particle trapping experiments performed to observe the effect of adjusting the oil menisci and the gap between two pairs of menisci in a four-menisci configuration on the nature of the DEP well formed at the center. A design parameter, Theta, capturing the dimensions of the DEP energy well, is defined and simulations exploring the effects of different geometric features on Theta are presented.     

Dielectrophoretic choking phenomenon of a deformable particle in a converging‐diverging microchannel

The translational motion of small particles in an electrokinetic fluid flow through a constriction can be enhanced by an increase of the applied electric potential. Beyond a critical potential, however, the negative dielectrophoresis (DEP) can overpower other forces to prevent particles that are even smaller than the constriction from passing through the constriction. This DEP choking phenomenon was studied previously for rigid particles. Here, the DEP choking phenomenon is revisited for deformable particles, which are ubiquitous in many biomedical applications. Particle deformability is measured by the particle shear modulus, and the choking conditions are reported through a parametric study that includes the channel geometry, external electric potential, and particle zeta potential. The study was carried out using a numerical model based on an arbitrary Lagrangian‐Eulerican (ALE) finite‐element method.     

Dielectrophoretic deformation of breast cancer cells for lab on a chip applications

This paper presents the development and experimental analysis of a curved microelectrode platform for the DEP deformation of breast cancer cells (MDA‐MB‐231). The platform is composed of arrays of curved DEP microelectrodes which are patterned onto a glass slide and samples containing MDA‐MB‐231 cells are pipetted onto the platform's surface. Finite element method is utilised to characterise the electric field gradient and DEP field. The performance of the system is assessed with MDA‐MB‐231 cells in a low conductivity 1% DMEM suspending medium. We applied sinusoidal wave AC potential at peak to peak voltages of 2, 5, and 10 V_pp at both 10 kHz and 50 MHz. We observed cell blebbing and cell shrinkage and analyzed the percentage of shrinkage of the cells. The experiments demonstrated higher percentage of cell shrinkage when cells are exposed to higher frequency and peak to peak voltage electric field.     

Positional dependence of particles in microfludic impedance cytometry

Single cell impedance cytometry is a label-free electrical analysis method that requires minimal sample preparation and has been used to count and discriminate cells on the basis of their impedance properties. This paper shows experimental and numerically simulated impedance signals for test particles (6 μm diameter polystyrene) flowing through a microfluidic channel. The variation of impedance signal with particle position is mapped using numerical simulation and these results match closely with experimental data. We demonstrate that for a nominal 40 μm × 40 μm channel, the impedance signal is independent of position over the majority of the channel area, but shows large experimentally verifiable variation at extreme positions. The parabolic flow profile in the channel ensures that most of the sample flows through the area of uniform signal. At high flow rates inertial focusing is observed; the particles flow in equal numbers through two equilibrium positions reducing the coefficient of variance (CV) in the impedance signals to negligible values.     

Lateral and cross-lateral focusing of spherical particles in a square microchannel

The inertial migration of particles in micro-scale flows has received much attention due to its promising applications, such as the membrane-free passive separation of particles or cells. The particles suspended in rectangular channels are known to be focused near the center of each channel face as the channel Reynolds number (R(C)) increases due to the lift force balance and the hydrodynamic interactions of the particles with the wall. In this study, the three-dimensional positions of neutrally buoyant spherical particles inside a square microchannel are measured using the digital holographic microscopy technique, and a transition from the lateral tubular pinch to the cross-lateral focusing with increasing R(C) is reported. The particles are found to migrate first in the lateral direction and then cross-laterally toward the four equilibrium positions. A general criterion that can be used to secure the fully developed state of particle focusing in Lab-on-a-Chip applications is also derived. This criterion could be helpful for the accurate estimation of the design parameters of inertial microfluidic devices, such as R(C), channel length and width, and particle diameter.     

Dielectrophoretic investigation of plant virus particles: Cow Pea Mosaic Virus and Tobacco Mosaic Virus

This paper reports experimental results on the dielectrophoretic (DEP) behaviour on two nonenveloped plant viruses of different geometrical shapes, namely Cow Pea Mosaic Virus (CPMV) and Tobacco Mosaic Virus (TMV). The DEP properties of carboxy-modified latex beads of the same size are also reported. The DEP properties of single particles were obtained from measurement of the frequency at which the DEP force on a particle goes to zero (the crossover frequency). The DEP behaviour of particle ensembles was also measured using image processing. The dielectric properties of the particles were evaluated from the DEP data. The surface conductance was found to be 0.3 nS for CPMV, 0.38 nS for TMV, and 0.52 nS for 27 nm diameter carboxy-latex beads. Data analysis has shown that the optimal condition for separation of TMV and CPMV is a low-conductivity suspending medium - below 1 mS/m.