Electrokinetic Focusing Research Articles

Electrokinetic focusing and separation of mammalian cells in conductive biological fluids.

Active manipulation of cells, such as trapping, focusing, and isolation, is essential for various bioanalytical applications. Herein, we report a hybrid electrokinetic technique for manipulating mammalian cells in physiological fluids. This technique applies a combination of negative dielectrophoretic force and hydrodynamic drag force induced by electrohydrodynamics, which is effective in conductive biological fluids. With a three-electrode configuration, the stable equilibrium positions of cells can be adjusted for separation and focusing applications. Cancer cells and white blood cells can be positioned and isolated into specific locations in the microchannel under both static and dynamic flow conditions. To investigate the sensitivity of the hybrid electrokinetic process, AC voltage, frequency, and bias dependences of the cell velocity were studied systematically. The applicability of the hybrid electrokinetic technique for manipulating cells in physiological samples is demonstrated by continuous focusing of human breast adenocarcinoma spiked in urine, buffy coats, and processed blood samples with 98% capture efficiency.

A low-cost 2D fluorescence detection system for μm sized beads on-chip

In this paper we describe a compact fluorescence detection system for on-chip analysis of beads, comprising a low-cost optical HD-DVD pickup. The complete system consists of a fluorescence detection unit, a control unit and a microfluidic chip containing microchannels and optical markers. With these markers the laser beam of the optical pickup can be automatically focused at the centre of the microchannel. With the complete system a two-dimensional fluorescent profile across the channel width can be obtained such that there is no need for hydrodynamic or electrokinetic focusing of the particles in a specific part of the channel. Fluorescent μm sized beads suspended in medium have been detected with the system. Since on both sides of the main beam two additional laser beams at a known distance are generated, also the velocity of individual beads has been determined.

High-throughput electrokinetic bioparticle focusing based on a travelling-wave dielectrophoretic field

This study presents a sheathless and portable microfluidic chip that is capable of high-throughput focusing bioparticles based on 3D travelling-wave dielectrophoresis (twDEP). High-throughput focusing is achieved by sustaining a centralized twDEP field normal to the continuous through-flow direction. Two twDEP electrode arrays are formed from upper and lower walls of the microchannel and extend to its center, which induce twDEP forces to provide the lateral displacements in two directions for focusing the bioparticles. Bioparticles can be focused to the center of the microchannel effectively by twDEP conveyance when the characteristic time due to twDEP conveying in the y direction is shorter than the residence time of the particles within twDEP electrode array. Red blood cells can be effectively focused into a narrow particle stream (~10 μm) below a critical flow rate of 10 μl/min (linear flow velocity ~5 mm/s), when under a voltage of 14 Vp–p at a frequency of 500 kHz is applied. Approximately 90% focusing efficiency for red blood cells can be achieved within two 6-mm-long electrode arrays when the flow rate is below 12 μl/min. Blood cells and Candida cells were also focused and sorted successfully based on their different twDEP mobilities. Compared to conventional 3D-paired DEP focusing, velocity is enhanced nearly four folds of magnitude. 3D twDEP provides the lateral displacements of particles and long residence time for migrating particles in a high-speed continuous flow, which breaks through the limitation of many electrokinetic cell manipulation techniques.

Confocal Fluctuation Spectroscopy and Imaging

Currently, work with subnanomolar concentrations is routine while femtomolar and even single-molecule studies are possible with some efforts getting high on single-molecule biophysics and biochemistry. Methodological breakthroughs, such as reducing the background light contribution in single-molecule studies, which has plagued many studies of molecular fluorescence in dilute solution, and particularly in live cells, have recently described by us. We first demonstrated how optimized time-gating of the fluorescence signal, together with time-correlated, single-photon counting, can be used to substantially boost the experimental signal-to-noise ratio about 140-fold, making it possible to measure analyte concentrations that are as low as 15 pM. By detection of femtomolar bulk concentrations, confocal microsopy has the potential to address the observation of one and the same molecule in dilute solution without immobilization or hydrodynamic/electrokinetic focusing at longer observation times than currently available. We present relevant physics. The equations are derived using Einstein's approach showing how it fits with Fick's law and the autocorrelation function. An improved technology is being developed at ISS for femtomolar microscopy. The general concepts and provided experimental examples should help to compare our approach to those used in conventional confocal microscopy.

Three-dimensional electrokinetic particle focusing in a rectangular microchannel

We extend an earlier study (Liang et al., 2010 [1]) to demonstrate a three-dimensional focusing of neutrally buoyant particles in electrophoresis through a rectangular microchannel as a result of the wall-induced lateral migration.

Electrokinetic focusing and filtration of cells in a serpentine microchannel.

Focusing cells into a single stream is usually a necessary step prior to counting and separating them in microfluidic devices such as flow cytometers and cell sorters. This work presents a sheathless electrokinetic focusing of yeast cells in a planar serpentine microchannel using dc-biased ac electric fields. The concurrent pumping and focusing of yeast cells arise from the dc electrokinetic transport and the turn-induced acdc dielectrophoretic motion, respectively. The effects of electric field (including ac to dc field ratio and ac field frequency) and concentration (including buffer concentration and cell concentration) on the cell focusing performance were studied experimentally and numerically. A continuous electrokinetic filtration of E. coli cells from yeast cells was also demonstrated via their differential electrokinetic focusing in a serpentine microchannel.

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

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

Dual‐asymmetry electrokinetic flow focusing for pre‐concentration and analysis of catecholamines in CE electrochemical nanochannels

In this research, a technique incorporating dual-asymmetry electrokinetic flow (DAEKF) was applied to a nanoCE electrochemical device for the pre-concentration and detection of catecholamines. The DAEKF was constructed by first generating a zeta-potential difference between the top and bottom walls, which had been pre-treated with O2 and H2O surface plasma, respectively, yielding a 2-D gradient shear flow across the channel depth. The shear flow was then exposed to a varying zeta-potential along the downstream direction by control of the field-effect in order to cause downward rotational flow in the channel. By this mechanism, almost all of the samples were effectively brought down to the electrode surface for analysis. Simulations were carried out to reveal the mechanism of concentration caused by the DAEKF, and the results reasonably describe our experiment findings. This DAEKF technique was applied to a glass/glass CE electrochemical nanochip for the analysis of catecholamines. The optimum detection limit was determined to be 1.25 and 3.3 nM of dopamine and catechol, respectively. A detection limit at the zeptomole level for dopamine can be obtained in this device, which is close to the level released by a single neuron cell in vitro.

Integrated Electrokinetic Sample Focusing and Surface Plasmon Resonance Imaging System for Measuring Biomolecular Interactions

Label-free biomolecular binding measurement methods, such as surface plasmon resonance (SPR), are becoming increasingly more important for the estimation of real-time binding kinetics. Recent advances in surface plasmon resonance imaging (iSPR) are emerging for label-free microarray-based assay applications, where multiple biomolecular interactions can be measured simultaneously. However, conventional iSPR microarray systems rely on protein printing techniques for ligand immobilization to the gold imaging surface and external pumps for analyte transport. In this article, we present an integrated microfluidics and iSPR platform that uses only electrokinetic transport and guiding of ligands and analytes and, therefore, requires only electrical inputs for sample transport. An important advantage of this new approach, compared to conventional systems, is the ability to direct a single analyte to a specific ligand location in the microarray, which can facilitate analysis parallelization. Additionally, this simple approach does not require complicated microfluidic channel arrangements, external pumps, or valves. As a demonstration, kinetics and affinity have been extracted from measured binding responses of human IgG and goat antihuman IgG using a simple 1:1 model and compared to responses measured with conventional pressure driven analyte transport. The measured results indicate similar binding kinetics and affinity between the electrokinetic and pressure-driven sample manipulation methods and no cross contamination to adjacent measurement locations has been observed.

Numerical and experimental evaluation of microfluidic sorting devices

The development of lab-on-a-chip devices calls for the isolation or separation of specific bioparticles or cells. The design of a miniaturized cell-sorting device for handheld operation must follow the strict parameters associated with lab-on-a-chip technology. The limitations include applied voltage, high efficiency of cell-separation, reliability, size, flow control, and cost, among others. Currently used designs have achieved successful levels of cell isolation; however, further improvements in the microfluidic chip design are important to incorporate into larger systems. This study evaluates specific design modifications that contribute to the reduction of required applied potential aiming for developing portable devices, improved operation reliability by minimizing induced pressure disturbance when electrokinetic pumping is employed, and improved flow control by incorporating directing streams achieving dynamic sorting and counting. The chip designs fabricated in glass and polymeric materials include asymmetric channel widths for sample focusing, nonuniform channel depth for minimizing induced pressure disturbance, directing streams to assist particle flow control, and online filters for reducing channel blockage. Fluorescence-based visualization experimental results of electrokinetic focusing, flow field phenomena, and dynamic sorting demonstrate the advantages of the chip design. Numerical simulations in COMSOL are validated by the experimental data and used to investigate the effects of channel geometry and fluid properties on the flow field.

New advances in on-line sample preconcentration by capillary electrophoresis using dynamic pH junction

The small injection volumes and narrow dimensions characteristic of microseparation techniques place constraints on concentration sensitivity that is required for trace chemical analyses. On-line sample preconcentration techniques using dynamic pH junction and its variants have emerged as simple yet effective strategies for enhancing concentration sensitivity of weakly ionic species by capillary electrophoresis (CE). Dynamic pH junction offers a convenient format for electrokinetic focusing of dilute sample plugs directly in-capillary for improved detection without off-line sample pretreatment. In this report, we highlight new advances in dynamic pH junction which have been reported to enhance method performance while discussing challenges for future research.

A microfluidic device for array patterning by perpendicular electrokinetic focusing

This paper describes a microfluidic chip in which two perpendicular laminar-flow streams can be operated to sequentially address the surface of a flow-chamber with semi-parallel sample streams. The sample streams can be controlled in position and width by the method of electrokinetic focusing. For this purpose, each of the two streams is sandwiched by two parallel sheath flow streams containing just a buffer solution. The streams are being electroosmotically pumped, allowing a simple chip design and a setup with no moving parts. Positioning of the streams was adjusted in real-time by controlling the applied voltages according to an analytical model. The perpendicular focusing gives rise to overlapping regions, which, by combinatorial (bio) chemistry, might be used for fabrication of spot arrays of immobilized proteins and other biomolecules. Since the patterning procedure is done in a closed, liquid filled flow-structure, array spots will never be exposed to air and are prevented from drying. With this device configuration, it was possible to visualize an array of 49 spots on a surface area of 1 mm2. This article describes the principle, fabrication, experimental results, analytical modeling and numerical simulations of the microfluidic chip.

Fabrication of UV epoxy resin masters for the replication of PDMS-based microchips

This paper presents two methods for the fabrication of UV epoxy resin masters for the replication of PDMS-based microfluidic chips. In the first method, the epoxy resin master is fabricated from a negative glass template manufactured using conventional lithography and wet etching techniques. However, in the second method, the master is produced simply by exposing a layer of UV epoxy resin coated on a glass substrate. Although the first method enables the replication of multiple PDMS structures from a single master, the latter method avoids the requirement for a wet chemical etching process and enables the epoxy master to be produced in 40 min or less. The experimental results show that the epoxy resin masters enable the mass production of PDMS replicas with highly precise geometrical tolerances. A series of electrokinetic focusing experiments are performed using PDMS microchips replicated from the current epoxy resin masters. The experimental results obtained for the width of the electrokinetically-focused sample stream under different focusing ratios are found to be in good agreement with the theoretical predictions. The sample handling characteristics of the microfluidic chips are also investigated. It is shown that the sample flow can be electrokinetically pre-focused into a narrow stream and then guided to the desired outlet port by applying a simple voltage control model. Finally, it is demonstrated that through an appropriate alignment of the sample flow and the conductivity gradient, the electrokinetic instability phenomenon can be induced at a relatively low electrical field strength of 0.35 kV/cm.

Electrokinetic flow focusing and valveless switching integrated with electrokinetic instability for mixing enhancement

This study performs an experimental investigation into electrokinetically-driven flow phenomena in microfluidic chips. The study commences by investigating the electrokinetic focusing/valveless switching of multiple sample flows in an M × N microfluidic chip, where M is the number of sample streams and N is the number of outlet channels. The experimental results show that the sample flows can be electrokinetically pre-focused into narrow streams via sheath flows and then guided to the desired outlet ports by applying a simple voltage control model which specifies the intensity of the external electrical field to be applied to the inlet channels and the required potential conditions of the individual outlet channels, i.e. grounded or isolated. The study then investigates the electrokinetic instability phenomenon induced at the interfaces between the sample flows and the sheath flows by the application of an external electrical field of sufficient intensity. It is shown that inducing this instability phenomenon yields a significant improvement in the mixing performance within the microchannel. Finally, it is demonstrated that by applying a high electrical potential to the inlet channel and specifying appropriate potential conditions for the individual outlet channels, two sample streams can be directed to any one of the N outlet channels in order to carry out mixing via electrokinetic instability effects.

Numerical analysis of an electrokinetic double‐focusing injection technique for microchip CE

The injection techniques in electrophoresis microchips play an important role in the sample-handling process, whose characteristics determine the separation performance achieved, and the shape of a sample plug delivered into the separation channel has a great impact on the high-quality separation performance as well. This paper describes a numerical investigation of different electrokinetic injection techniques to deliver a sample plug within electrophoresis microchips. A novel double-focusing injection system is designed and fabricated, which involves four accessory arm channels in which symmetrical focusing potentials are loaded to form a unique parallel electric field distribution in the intersection of injection channel and separation channel. The parallel electric field effectuates virtual walls to confine the spreading of a sample plug at the intersection and prevents sample leakage into separation channel during the dispensing step. The key features of this technique over other injection techniques are the abilities to generate regular and nondistorted shape of sample plugs and deliver the variable-volume sample plugs by electrokinetic focusing. The detection peak in the proposed injection system is uniform regardless of the position of the detection probe in the separation channel, and the peak resolution is greatly enhanced. Finally, the double-focusing injection technique shows the flexibility in detection position and ensures improved signal sensitivity with good peak resolution due to the delivered high-quality sample plug.

Convective instability of electrokinetic flows in a cross-shaped microchannel

We present a parametric experimental study of convective electrokinetic instability (EKI) in an isotropically etched, cross-shaped microchannel using quantitative epifluorescence imaging. The base state is a three-inlet, one-outlet electrokinetic focusing flow configuration where the centre sample stream and sheath flows have mismatched ionic conductivities. Electrokinetic flows with conductivity gradients become unstable when the electroviscous stretching and folding of conductivity interfaces grows faster than the dissipative effect of molecular diffusion. Scalar images, critical applied fields required for instability, and temporal and spatial scalar energy are presented for flows with a wide range of applied d.c. electric field and centre-tosheath conductivity ratios. These parameters impose variations of the electric Rayleigh number across four orders of magnitude. We introduce a scaling for charge density in the bulk fluid as a function of local maximum conductivity gradients in the flow. This scaling shows that the flow becomes unstable at a critical electric Rayleigh number (Ra e,l =205) and applies to a wide range of applied field and centre-to-sheath conductivity ratios. This work is relevant to on-chip electrokinetic flows with conductivity gradients such as field amplified sample stacking, flow at the intersections of multi-dimensional assays, electrokinetic control and separation of sample streams with poorly specified chemistry, and low-Reynolds number micromixing.

Numerical modeling of the Joule heating effect on electrokinetic flow focusing

In electrokinetically driven microfluidic systems, the driving voltage applied during operation tends to induce a Joule heating effect in the buffer solution. This heat source alters the solution's characteristics and changes both the electrical potential field and the velocity field during the transport process. This study performs a series of numerical simulations to investigate the Joule heating effect and analyzes its influence on the electrokinetic focusing performance. The results indicate that the Joule heating effect causes the diffusion coefficient of the sample to increase, the potential distribution to change, and the flow velocity field to adopt a nonuniform profile. These variations are particularly pronounced under tighter focusing conditions and at higher applied electrical intensities. In numerical investigations, it is found that the focused bandwidth broadens because thermal diffusion effect is enhanced by Joule heating. The variation in the potential distribution induces a nonuniform flow field and causes the focused bandwidth to tighten and broaden alternately as a result of the convex and concave velocity flow profiles, respectively. The present results confirm that the Joule heating effect exerts a considerable influence on the electrokinetic focusing ratio.

A new focusing model and switching approach for electrokinetic flow inside microchannels

This paper presents an investigation into two crucial aspects of microfluidic applications, namely electrokinetic focusing and switching. This study commences by modeling the electrokinetic focusing phenomenon theoretically using the potential flow theory. A new theoretical model is derived and applied to predict the width of the focused stream. The results predicted by the theoretical model are shown to be in good agreement with the experimental data. The paper then proceeds to study the electrokinetic switching functions systematically using both experimental and theoretical approaches. A new control model for ‘one-to-multiple’ electrokinetically pre-focused micro flow switches is proposed. Using this new model, the sample flow can be pre-focused electrokinetically into a narrow stream and then injected directly into the desired outlet port. The results of this study provide a useful methodology for the analysis of flow control in microfluidic devices. Finally, an electrokinetically driven micro flow cytometer utilizing electrokinetic focusing and switching effects is demonstrated. Experimental data show that the developed methods could successfully focus microparticles and direct them into any desired outlet port.

Focused electrophoretic motion and selected electrokinetic dispensing of particles and cells in cross-microchannels

The electrokinetic focusing and the resultant accelerated electrophoretic motion of polystyrene particles and red blood cells were visualized in microfluidic cross-channels. The experimentally measured width of the focused stream and the measured velocity increase of particles and cells at different voltage ratios follow the proposed analytical formula within the experimental error. The attained velocity increase is insensitive to the particle size, particle property (i.e., particle or cell), and particle trajectory. By solving the electrical potential field in the cross-channel at the experimental conditions, we demonstrate that the squeezed electrical field lines in the channel intersection determine the shape of the focused stream, and the nonuniform distribution of axial electrical field strength underlies the variation of particle/cell electrophoretic velocity through the focusing region. However, the dielectrophoretic force resulting from the nonuniform electrical field in the intersection seems to push the acceleration region of particles and cells slightly in the downstream direction. We have also achieved the single particle/cell dispensing by instantly triggering an electrical pulse perpendicular to the focused particulate flow in a double-cross microchannel. The electrokinetic manipulation of particle/cell in microchannels demonstrated in this work can be used for developing integrated lab-on-a-chip devices for studies of cells.

Analytical Currents: New electrokinetic focusing method

ADVERTISEMENT RETURN TO ISSUEPREVNewsNEXTAnalytical Currents: New electrokinetic focusing methodCite this: Anal. Chem. 2004, 76, 9, 153 APublication Date (Web):May 1, 2004Publication History Published online1 May 2004Published inissue 1 May 2004https://doi.org/10.1021/ac041546xRIGHTS & PERMISSIONSArticle Views103Altmetric-Citations-LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (48 KB) Get e-AlertsSUBJECTS:Electrokinetics Get e-Alerts