Development of the resolution theory for electrophoretic exclusion

Electrophoretic exclusion, a technique that differentiates species in bulk solution near a channel entrance, has been demonstrated on benchtop and microdevice designs. In these systems, separation occurs when the electrophoretic velocity of one species is greater than the opposing hydrodynamic flow, while the velocity of the other species is less than that flow. Although exclusion has been demonstrated in multiple systems for a range of analytes, a theoretical assessment of resolution has not been addressed. To compare the results of these calculations to traditional techniques, the performance is expressed in terms of smallest difference in electrophoretic mobilities that can be completely separated (R = 1.5). The calculations indicate that closest resolvable species (Δμ_min) differ by approximately 10^?13 m²/Vs and peak capacity (n_c) is 1000. Published experimental data were compared to these calculated results.     

A study on the condition for differential electrophoretic transport at a channel entrance

Electrophoretic differential transport of ionic species in a solution moving from a large reservoir into a small channel is investigated numerically. The system setup is similar to the experiments of Polson, Savin, and Hayes (J. Microcol. Sep. 2000, 12, 98), where the bulk flow into a fused-silica capillary was driven by a pressure differential. A critical condition for achieving the defined differential transport near the channel entrance is found and this condition is solely determined by a dimensionless parameter when the geometry of the system is prescribed. This dimensionless parameter is the ratio between the electrophoretic migration velocity of the species based on the apparent electric intensity and the centerline fluid velocity of the fully developed channel flow. Species concentration distributions are also computed for various conditions. A separation technique can be derived from the experimental condition where a targeted division of species can be created at the channel entrance.     

Continuous microfluidic DNA and protein trapping and concentration by balancing transverse electrokinetic forces

Sample pre-concentration can be a critical element to improve sensitivity of integrated microchip assays. In this work a converging Y-inlet microfluidic channel with integrated coplanar electrodes was used to investigate transverse DNA and protein migration under uniform direct current (DC) electric fields to assess the ability to concentrate a sample prior to other enzymatic modifications or capillary electrophoretic separations. Employing a pressure-driven flow to perfuse the microchannel, negatively charged samples diluted in low and high ionic strength buffers were co-infused with a receiving buffer of the same ionic strength into a main daughter channel. Experimental results demonstrated that, depending of the buffer selection, different DNA migration and accumulation dynamics were seen. Charged analytes could traverse the channel width and accumulate at the positive bias electrode in a low electroosmotic mobility, high electrophoretic mobility, high ionic strength buffer or migrated towards an equilibrium position within the channel in a high electroosmotic mobility, high electrophoretic mobility, low ionic strength buffer. The various migration behaviours are the result of a balance between the electrophoretic force and a drag force induced by a recirculating electroosmotic flow generated across the channel width due to the bounding walls. Under continuous flow conditions, DNA samples were concentrated several-fold by balancing these transverse electrokinetic forces. The electrokinetic trapping technique presented here is a simple technique which could be expanded to concentrate or separate other analytes as a preconditioning step for downstream processes.     

Quantitative assessment of flow and electric fields for electrophoretic focusing at a converging channel entrance with interfacial electrode

The electric field and flow field gradients near an electrified converging channel are amenable to separating and focusing specific classes of electrokinetic material, but the detailed local electric field and flow dynamics in this region have not been thoroughly investigated. Finite elemental analysis was used to develop a model of a buffer reservoir connected to a smaller channel to simulate the electrophoretic and flow velocities (which correspond directly to the respective electric and flow fields) at a converging entrance. A detailed PTV (Particle Tracking Velocimetry) study using charged fluorescent microspheres was performed to assess the model validity both in the absence and presence of an applied electric field. The predicted flow velocity gradient from the model agreed with the PTV data when no electric field was present. Once the additional forces that act on the large particles required for tracing (dielectrophoresis) were included, the model accurately described the velocity of the charged particles in electric fields.     

Ultrafast electrokinetics

The influence of a high electric field applied on both fluid flow and particle velocities is quantified at large Peclet numbers. The experiments involved simultaneous particle image velocimetry and flow rate measurements. These are conducted in polydimethylsiloxane channels with spherical nonconducting polystyrene particles and DI water as the background flow. The high electric field tests produced up to three orders of magnitude higher electrokinetic velocities than any previous reports. The maximum electroosmotic velocity and electrophoretic velocity measured were 3.55 and 2.3 m/s. Electrophoretic velocities are measured over the range of 100 V/cm < E < 250 000 V/cm. The results are separated according to the different nonlinear theoretical models, including low and high Peclet numbers, and weak and strong concentration polarization. They show good agreement with the models. Such fast velocities could be used for flow separation, mixing, transport, control, and manipulation of suspended particles as well as microthrust generation among other applications.     

Bipolar electrode focusing: tuning the electric field gradient

Bipolar electrode (BPE) focusing is a developing technique for enrichment and separation of charged analytes in a microfluidic channel. The technique employs a bipolar electrode that initiates faradaic processes that subsequently lead to formation of an ion depletion zone. The electric field gradient resulting from this depletion zone focuses ions on the basis of their individual electrophoretic mobilities. The nature of the gradient is of primary importance to the performance of the technique. Here, we report dynamic measurements of the electric field gradient showing that it is stable over time and that its axial position in the microchannel is directly correlated to the location of an enriched tracer band. The position of the gradient can be tuned with pressure-driven flow. We also show that a steeper electric field gradient decreases the breadth of the enriched tracer band and therefore enhances the enrichment process. The slope of the gradient can be tuned by altering the buffer concentration: higher concentrations result in a steeper gradient. Coating the channel with the neutral block co-polymer Pluronic also results in enhanced enrichment.     

Electrically enhanced crossflow membrane filtration of oily waste water using the membrane as a cathode

The effect of an external electric field on the flux in crossflow membrane filtration of a model oily waste water was studied using a carbon fibre – carbon composite membrane as a cathode. Limiting fluxes for low flow rate increased significantly under the conditions studied, from 75 l/m² h without an electric field to more than 350 l/m² h using an electric field. The experimentally determined increase in the limiting flux showed good agreement with the theoretical value of 430 l/m² h calculated using a simple model. The limiting flux increase was affected by the electrophoretic mobility of the oil droplets and the applied electric field strength. When there were no cakes without an electric field due to the high flow rate, the flux increase when using an electric field under at the same conditions was minor. The critical electric field strength was determined, and experimentally obtained values were corresponded with calculated values. Decreasing the crossflow velocity above the critical electric field strength increased the flux, or had no effect, depending on the size of the particles. Permeate quality was also improved to some extent when using the electric field, and a membrane with a large pore size could be used when an electric field was applied. The main disadvantage in using the membrane as a cathode was foaming at the membrane surface causing decrease in the flux enhancement as the conductivity of the feed increased. It was not possible to restore the flux to the original value by applying an electric field after filtration of the oil emulsion without an electric field. An intermittent electric field was thus not efficient enough for keeping the flux at high level.     

Amperometric detection of amines and amino acids in flow injection systems with a nickel oxide electrode

The use and characteristics of a nickel oxide electrode as a detector for amines in a flow injection system are described. The anodic electrode reaction mechanism involves a higher oxidation state of nickel maintained by the applied potential (+0.49 V vs. SCE). The electroanalytical parameters are investigated and the currents for a series of amines and amino acids are compared. Two electrode configurations are compared. The flow injection technique is shown to be suitable for buffered 25-μl samples of pH as low as 3. The linear range for glycine is 10^-6–10^-3 M with detection limits of a few nanograms.     

Diatomics-in-molecules calculations of potential-energy surfaces for B+(3P) + H2(X 1Σg+)

Collisions of B⁺(³P) with H₂(X ¹Σ_g⁺) have been studied repeatedly using molecular-beam techniques. The theoretical interpretation of the results suffered from missing information about the potential-energy surfaces for this system. This paper reports on diatomics-in-molecules calculations for a wide range of nuclear geometries. The stationary points on the minimum-energy paths are determined. Symmetrically orthogonalized and non-hermitean diatomics-in-molecules versions differ only slightly at the minimum-energy path. The potential-energy surfaces of the 1³A′ and 2³A′ states show dramatical changes with the BHH angle, leading possibly to adiabatic as well as non-adiabatic elementary processes. The 1³A′' and 1³A′ states are nearly degenerate for geometries ranging from the entrance channel to the interaction region. The most favourable configurations of approach on these potential-energy surfaces are those with C₂, symmetry.     

Low-voltage driven control in electrophoresis microchips by traveling electric field.

This paper presents the use of a physical model and numerical simulation in the investigation of traveling electric fields on capillary electrophoresis (CE) chips. The principal material transport mechanisms of electrokinetic migration, ionic concentration, fluid flow, and diffusion are all taken into consideration. Traditionally, the high electric field strength required for the separation of biological samples by microfluidic devices has involved the application of high external voltages. In contrast, this study presents a proposal for samples separation by means of a moving electric field within a low voltage-driven CE chip. Under this proposal, the separation channel is partitioned into a series of smaller separation zones by means of electrode pairs. This paper considers two different electrode configurations, namely arranged along a single side of the separation channel, and arranged on two sides of the separation channel. The quality of the separation achieved with these two configurations is then compared with the traditional straight separation channel approach. The results confirm that the proposed method is successful in maintaining an adequate field strength for separation purposes in a low-voltage driven CE chip. Furthermore, it is determined that the best separation results are obtained using electrodes arranged along both sides of the separation channel.     

Using electrophoretic exclusion to manipulate small molecules and particles on a microdevice

Electrophoretic exclusion, a novel separations technique that differentiates species in bulk solution using the opposing forces of electrophoretic velocity and hydrodynamic flow, has been adapted to a microscale device. Proof-of-principle experiments indicate that the device was able to exclude small particles (1 μm polystyrene microspheres) and fluorescent dye molecules (rhodamine 123) from the entrance of a channel. Additionally, differentiation of the rhodamine 123 and polystyrene spheres was demonstrated. The current studies focus on the direct observation of the electrophoretic exclusion behavior on a microchip.     

Optimization of Field-Flow Fraction-Ation with Respect to Relaxation and Use of Stop-Flow Technique at Constant Field Operation

Abstract

The relaxation phenomena developing after the injection of the sample into the channel during Field-Flow Fractionation were analyzed theoretically. Relaxation times, necessary to stop the flow immediately after the injection, were calculated on the basis of various models describing the relaxation processes. A stop-flow time long enough so that the deviation from the equilibrium would not influence the retention within the limits of experimental errors and so that the maximum efficiency would be reached was required. Sedimentation Field-Flow Fractionation was used to verify the theoretical analysis. The diffusion coefficient of latex used for the study is small and thus the long relaxation times allow to reach sufficient precision of the measurement. The experiments performed at different flow rates and using different ways of sampling proved a good agreement between the experimental relaxation times and the values calculated theoretically. The stop-flow technique was proved both theoretically and experimentally to be advantageous from the viewpoint of shortening the time of fractionation The disagreement with the previous experimental data obtained by other authors was explained. There was a good agreement between our experimental data and the theoretical predictions. Field-Flow Fractionation using the stop-flow technique at the constant field operation was found to be very suitable for exact physico-chemical measurements.     

Performance optimization in electric field gradient focusing

Electric field gradient focusing (EFGF) is a technique used to simultaneously separate and concentrate biomacromolecules, such as proteins, based on the opposing forces of an electric field gradient and a hydrodynamic flow. Recently, we reported EFGF devices fabricated completely from copolymers functionalized with poly(ethylene glycol), which display excellent resistance to protein adsorption. However, the previous devices did not provide the predicted linear electric field gradient and stable current. To improve performance, Tris–HCl buffer that was previously doped in the hydrogel was replaced with a phosphate buffer containing a salt (i.e., potassium chloride, KCl) with high mobility ions. The new devices exhibited stable current, good reproducibility, and a linear electric field distribution in agreement with the shaped gradient region design due to improved ion transport in the hydrogel. The field gradient was calculated based on theory to be approximately 5.76 V/cm² for R-phycoerythrin when the applied voltage was 500 V. The effect of EFGF separation channel dimensions was also investigated; a narrower focused band was achieved in a smaller diameter channel. The relationship between the bandwidth and channel diameter is consistent with theory. Three model proteins were resolved in an EFGF channel of this design. The improved device demonstrated 14,000-fold concentration of a protein sample (from 2 ng/mL to 27 μg/mL).     

3-D transient electrophoretic motion of a spherical particle in a T-shaped rectangular microchannel

This paper considers the electrophoretic motion of a spherical particle in an aqueous electrolyte solution in a T-shaped rectangular microchannel, where the size of the channel is close to that of the particle. This is a complicated transient process where the electric field, the flow field, and the particle motion are coupled together. A theoretical model was developed to investigate the influences of the applied electric potentials, the zeta potentials of the channel and the particle, and the size of the particle on the particle motion. A direct numerical simulation method using the finite element method is employed. This method employs a generalized Galerkin finite element formulation that incorporates both equations of the fluid flow and equations of the particle motion into a single variational equation where the hydrodynamic interactions are eliminated. The ALE method is used to track the surface of the particle at each time step. The numerical results show that the electric field in the T-shaped microchannel is influenced by the presence of the particle, and that the particle motion is influenced by the applied electric potentials and the zeta potentials of the channel and the particle. The path of the particle motion is dominated by the local electric field and the ratio of the zeta potential of the channel to that of the particle. The particle's velocity is also dependent on its size in a small channel.     

Voltage-controlled separation of proteins by electromobility focusing in a dialysis hollow fiber

Electromobility focusing (EMF) is a relatively new protein separation technique that utilizes an electric field gradient and a hydrodynamic flow. Proteins are focused in order of electrophoretic mobility at points where their electrophoretic migration velocities balance the hydrodynamic flow velocity. Steady state bands are formed along the separation channel when equilibrium is reached. Further separation and detection can be easily achieved by changing the electric field profile. In this paper. we describe an EMF system with on-line UV absorption detection in which the electric field gradient was formed using a dialysis hollow fiber. Protein focusing and preconcentration were performed with this system. Voltage-controlled separation was demonstrated using bovine serum albumin and myoglobin as model proteins. The limitations of the current method are discussed, and possible solutions are proposed.     

微流控芯片中超微电极的制作及其在芯片-电化学检测中的应用

设计并制作了集成有超微电极的玻璃微流控芯片.在电化学检测芯片1(EC-1)中,以光刻方法制作13μm宽的Pt超微电极,距分离管道末端30μm,优化电极体系和分离电压,检测了电泳分离的神经递质.在电化学检测芯片2(EC-2)中,制作7μm宽的超微电极,在其上游集成城墙式的膜结构,进一步腐蚀后的膜厚度为10μm,具有良好的导电性和散热性能,成功地将高压电场截至在超微电极之前,具有进一步应用于电化学检测的能力.     

Sequential injection with lab-at-valve (LAV) approach for potentiometric determination of chloride

Sequential injection with “Lab-at-Valve (LAV)” approach is demonstrated for potentiometric determination of chloride. The LAV flow-through electrode system consists of two Ag/AgCl electrodes: one as a reference electrode, silver chloride activated surface-silver wire soaked in a constant-concentration chloride ion solution in a small tube covered with a polymer-membrane, another as a working electrode (a similar silver chloride activated surface-silver wire) placed in a flow channel. The electrode system is attached at one port of a 10 port multiposition valve. A modified autoburette was used as a propelling device. Using SI operation via a program written in-house, based on LabVIEW^®, a standard/sample is inserted, via the selection valve, in potassium nitrate as an electrolyte and water is used as a carrier. The zones are transported from the holding coil to the flow cell to monitor the difference in potential due to concentration cell behavior. The potential difference is then recorded as a peak. Peak height is proportional to logarithm of chloride concentration. The SI-LAV for chloride determination is very simple, fast, precise, accurate, automatic and economical. Applications to mineral drinking water and surface water have been made. The results agree with those of IC and titrimetric methods.     

Bi-directional flow induced by an AC electroosmotic micropump with DC voltage bias

This paper discusses the principle of biased alternating current electroosmosis (ACEO) and its application to move the bulk fluid in a microchannel, as an alternative to mechanical pumping methods. Previous EO-driven flow research has looked at the effect of electrode asymmetry and transverse traveling wave forms on the performance of electroosmotic pumps. This paper presents an analysis that was conducted to assess the effect of combining an AC signal with a DC (direct current) bias when generating the electric field needed to impart electroosmosis (EO) within a microchannel. The results presented here are numerical and experimental. The numerical results were generated through simulations performed using COMSOL 3.5a. Currently available theoretical models for EO flows were embedded in the software and solved numerically to evaluate the effects of channel geometry, frequency of excitation, electrode array geometry, and AC signal with a DC bias on the flow imparted on an electrically conducting fluid. Simulations of the ACEO flow driven by a constant magnitude of AC voltage over symmetric electrodes did not indicate relevant net flows. However, superimposing a DC signal over the AC signal on the same symmetric electrode array leads to a noticeable net forward flow. Moreover, changing the polarity of electrical signal creates a bi-directional flow on symmetrical electrode array. Experimental flow measurements were performed on several electrode array configurations. The mismatch between the numerical and experimental results revealed the limitations of the currently available models for the biased EO. However, they confirm that using a symmetric electrode array excited by an AC signal with a DC bias leads to a significant improvement in flow rates in comparison to the flow rates obtained in an asymmetric electrode array configuration excited just with an AC signal.     

Controlled protein adsorption on a silica microparticle

In the present study, controlled protein adsorption on a rigid silica microparticle is investigated numerically using classical Langmuir and two-state models under electrokinetic flow conditions. The instantaneous particle locations are simulated along a straight microchannel using an arbitrary Lagrangian−Eulerian framework in the finite element method for the electrophoretic motion of the charged particle. Within the scope of the parametric study, the strength of the external electric field (E), particle diameter (D_p), the zeta potential of the particle (ζ_p), and the location of the microparticle away from the channel wall (H) are systematically varied. The results are also compared to the data of pressure-driven flow having a parabolic flow profile at the inlet whose maximum magnitude is set to the particle's electrophoretic velocity magnitude. The validation studies reveal that the code developed for the particle motion in the present simulations agrees well with the experimental results. It is observed that protein adsorption can be controlled using electrokinetic phenomena. The plug-like flow profile in electrokinetics is beneficial for a microparticle at every spatial location in the microchannel, whereas it is not valid for the pressure-driven flow. The electric field strength and the zeta potential of the particle accelerate the protein adsorption. The wall shear stress and shear rate are good indicators to predict the adsorption process for electrokinetic flow.