Performance impact of dynamic surface coatings on polymeric insulator-based dielectrophoretic particle separators.

Efficient and robust particle separation and enrichment techniques are critical for a diverse range of lab-on-a-chip analytical devices including pathogen detection, sample preparation, high-throughput particle sorting, and biomedical diagnostics. Previously, using insulator-based dielectrophoresis (iDEP) in microfluidic glass devices, we demonstrated simultaneous particle separation and concentration of various biological organisms, polymer microbeads, and viruses. As an alternative to glass, we evaluate the performance of similar iDEP structures produced in polymer-based microfluidic devices. There are numerous processing and operational advantages that motivate our transition to polymers such as the availability of numerous innate chemical compositions for tailoring performance, mechanical robustness, economy of scale, and ease of thermoforming and mass manufacturing. The polymer chips we have evaluated are fabricated through an injection molding process of the commercially available cyclic olefin copolymer Zeonor 1060R. This publication is the first to demonstrate insulator-based dielectrophoretic biological particle differentiation in a polymeric device injection molded from a silicon master. The results demonstrate that the polymer devices achieve the same performance metrics as glass devices. We also demonstrate an effective means of enhancing performance of these microsystems in terms of system power demand through the use of a dynamic surface coating. We demonstrate that the commercially available nonionic block copolymer surfactant, Pluronic F127, has a strong interaction with the cyclic olefin copolymer at very low concentrations, positively impacting performance by decreasing the electric field necessary to achieve particle trapping by an order of magnitude. The presence of this dynamic surface coating, therefore, lowers the power required to operate such devices and minimizes Joule heating. The results of this study demonstrate that iDEP polymeric microfluidic devices with surfactant coatings provide an affordable engineering strategy for selective particle enrichment and sorting.

Fabrication and analysis of spatially uniform field electrokinetic flow devices: theory and experiment.

A uniform-field design approach can improve the performance of microanalytical, chip-based devices for a number of applications, including separations and sample preparation. The faceted prism paradigm allows the design of microfluidic devices possessing spatially uniform fields in electrokinetically driven flows. We present the first quantitative study of the velocity fields obtained using faceted interfaces between deep and shallow channel sections. Electrokinetic flows were generated in a series of wet-etch fabricated microfluidic channels. The resulting velocity fields were analyzed by particle image velocimetry and compared with simulations of the two-dimensional Laplace equation using both the designed channel geometry and the as-fabricated channel geometry. This analysis found localized differences between the designed and observed flow fields that were directly attributable to the limitations of isotropic substrate etching. Simulations using the as-fabricated channel geometry reproduced the experimental electrokinetic velocity field, quantitatively accounting for speed field variations due to the limits of the fabrication method. The electrokinetic speed fields were also compared to corresponding pressure-driven speed fields.

Dielectrophoretic manipulation of particles and cells using insulating ridges in faceted prism microchannels.

This paper presents a novel device for the dielectrophoretic manipulation of particles and cells. A two-level isotropic etch of a glass substrate was used to create three-dimensional ridge-like structures in micrometer-sized channels. Due to the insulating properties of glass, locally patterned regions of nonuniform electric field form near the ridges when a dc field is applied along the channel. The ridges are designed using the method of faceted prisms, such that substantially uniform fields are produced on each side of the faceted interfaces that form each ridge. The dielectrophoretic force that results from the electric field gradient near the ridges is used to affect particle motion parallel to the ridges in the absence of a bulk pressure-driven flow. Trapping and deflection of particles and continuous concentration and separation of Bacillus subtilis from a two-component sample mixture are demonstrated. The flow of B. subtilis is restricted to a selected channel of a planar, multichannel device as a result of negative dielectrophoresis arising from the presence of the insulating ridges when the applied electric field exceeds a threshold of 30 V/mm. Dielectrophoresis has a negligible impact on 200-nm-diameter polystyrene particles under the same conditions.

An insulator-based (electrodeless) dielectrophoretic concentrator for microbes in water.

Dielectrophoresis (DEP), the motion of a particle caused by an applied electric field gradient, can concentrate microorganisms non-destructively. In insulator-based dielectrophoresis (iDEP) insulating microstructures produce non-uniform electric fields to drive DEP in microsystems. This article describes the performance of an iDEP device in removing and concentrating bacterial cells, spores and viruses while operated with a DC applied electric field and pressure gradient. Such a device can selectively trap particles when dielectrophoresis overcomes electrokinesis or advection. The dielectrophoretic trapping behavior of labeled microorganisms in a glass-etched iDEP device was observed over a wide range of DC applied electric fields. When fields higher than a particle-specific threshold are applied, particles are reversibly trapped in the device. Experiments with Bacillus subtilis spores and the Tobacco Mosaic Virus (TMV) exhibited higher trapping thresholds than those of bacterial cells. The iDEP device was characterized in terms of concentration factor and removal efficiency. Under the experimental conditions used in this study with an initial dilution of 1 x 105 cells/ml, concentration factors of the order of 3000x and removal efficiencies approaching 100% were observed with Escherichia coli cells. These results are the first characterization of an iDEP device for the concentration and removal of microbes in water.

The zeta potential of cyclo-olefin polymer microchannels and its effects on insulative (electrodeless) dielectrophoresis particle trapping devices.

While cyclo-olefin polymer microchannels have the potential to improve both the optical detection sensitivity and the chemical resistance of polymer microanalytical systems, their surface properties are to date not thoroughly characterized. These surface properties dictate, among other things, electrokinetic effects when electric fields are present. Here, we report the measurement of the zeta potential of cyclo-olefin polymers (injection-molded and hot-embossed Zeonor 1060R and 1020R) microchannels as a function of pH, counter-ion concentration, storage conditions, and chemical treatment in aqueous solutions both with and without EOF-suppressing additives. In contrast with previous reports, significant surface charge is measured, consistent with titration of charged sites with pK(a) = 4.8. Storage in air, acetonitrile, or aqueous solutions has relatively minor effects. While the source of the surface charge is unclear, chemical functionalization has shown that carboxylic acid groups are not present at the surface, consistent with the chemical structure of Zeonor. EOF-suppressing additives (hydroxypropylmethylcellulose) and conditioning in perchloric acid allow the surface charge to be suppressed. We demonstrate dielectrophoretic particle trapping devices in Zeonor 1060R substrates that show reduced trapping voltage thresholds as compared to previous implementations in glass.

Insulator-based dielectrophoresis for the selective concentration and separation of live bacteria in water.

Insulator-based dielectrophoresis (iDEP) was utilized to separate and concentrate selectively mixtures of two species of live bacteria simultaneously. Four species of bacteria were studied: the Gram-negative Escherichia coli and the Gram-positive Bacillus subtilis, B. cereus, and B. megaterium. Under an applied direct current (DC) electric field all the bacterial species exhibited negative dielectrophoretic behavior. The dielectrophoretic separations were carried out in a glass microchannel containing an array of insulating posts. The insulating posts in the microchannel produced nonuniformities in the electric field applied along the channel. Mixtures of two species of bacteria were introduced into the microchannel and the electric field was applied. The bacterial species exhibited different dielectrophoretic mobilities under the influence of the nonuniform field. From these experiments a trapping order was established with E. coli trapping at the weakest applied electric field, while the Bacillus species were trapped at different characteristic threshold fields. At stronger applied electric fields, the two different species of bacteria in the microchannel were dielectrophoretically trapped into two spatially distinct bands. The results showed that iDEP has the potential to selectively concentrate and separate different species of bacteria.

Dielectrophoretic concentration and separation of live and dead bacteria in an array of insulators.

Insulator-based (electrodeless) dielectrophoresis (iDEP) is an innovative approach in which the nonuniform electric field needed to drive DEP is produced by insulators, avoiding problems associated with the use of electrodes. Live and dead Escherichia coli were concentrated and selectively released by applying stepped DC voltages across a microchannel containing an array of insulating posts etched in glass. The only electrodes present were two platinum wires placed in the inlet and outlet reservoirs, producing mean electric fields of up to 200 V/mm across the insulators. The cells were labeled with Syto 9 and propidium iodide and imaged through a fluorescent microscope. Cell trapping and release were controlled by modifying the relative responses of electrokinesis and DEP by adjusting the magnitude of the applied voltage. Dead cells were observed to have significantly lower dielectrophoretic mobility than live cells, whereas the electrokinetic mobilities of live and dead cells were indistinguishable. The locations of the bands of differentially trapped cells were consistent with predictions. In addition, cells were selectively trapped and concentrated against backgrounds of 1- and 0.2-microm carboxylate-modified polystyrene particles. This first application of iDEP for simultaneous live/dead bacteria separation and concentration illustrates its potential as a front-end method for bacterial analysis.

Low-dispersion electrokinetic flows for expanded separation channels in microfluidic systems: multiple faceted interfaces.

A novel methodology to design on-chip conduction channels is presented for expansion of low-dispersion separation channels. Designs are examined using two-dimensional numerical solutions of the Laplace equation with a Monte Carlo technique to model diffusion. The design technique relies on trigonometric relations that apply for ideal electrokinetic flows. Flows are rotated and stretched along the abrupt interface between adjacent regions having differing specific permeability. Multiple interfaces can be placed in series along a channel. The resulting channels can be expanded to extreme widths while minimizing dispersion of injected analyte bands. These channels can provide a long path length for line-of-sight optical absorption measurements. Expanded sections can be reduced to enable point detection at the exit section of the channel. Designed to be shallow, these channels have extreme aspect ratios in the wide section, greatly increasing the surface-to-volume ratio to increase heat removal and decrease unwanted pressure-driven flow. The use of multiple interfaces is demonstrated by considering several three-interface designs. Faceted flow splitters can be constructed to divide channels into any number of exit channels while minimizing dispersion. The resulting manifolds can be used to construct medians for structural support in wide, shallow channels.

Dielectrophoresis in microchips containing arrays of insulating posts: theoretical and experimental results.

Dielectrophoresis (DEP), a nonlinear electrokinetic transport mechanism, can be used to concentrate and sort cells, viruses, and particles. To date, microfabricated DEP-based devices have typically used embedded metal electrodes to apply spatially nonuniform, time-varying (AC) electric fields. We have developed an alternative method in which arrays of insulating posts in a channel of a microchip produce the spatially nonuniform fields needed for DEP. Electrodes may be located remotely, allowing operation of the device down to zero frequency (DC) without excessive problems of electrolysis. Applying a sufficiently large electric field across an insulating-post array produces two flow regimes through a competition between electrokinetic flow (combined electrophoresis and electroosmosis) and dielectrophoresis. "Streaming DEP" is observed when DEP dominates diffusion but is overcome by electrokinetic flow. Particles concentrated by DEP forces in areas of electric field extrema travel electrokinetically down the array in flowing streams. In an array of posts, dielectrophoretic forcing within repeated rows adds coherently to produce flowing streams of highly concentrated and rarefied particles. We demonstrate that this reinforcement is a strong function of alignment of the array with respect to the applied electric field and that the particle concentrations can be "enhanced" or "depleted" along columns of posts, enabling a novel class of continuous-flow, selective particle filter/concentrator devices. To our knowledge, this is the first observation of streaming dielectrophoresis. The second regime is "trapping DEP," in which DEP forces dominate over both diffusion and electrokinetic flow, reversibly immobilizing particles on the insulating posts, enabling inexpensive and embedded batch filter/concentrator devices. Devices can be biased electrically to manipulate particles selectively by varying the field strength to vary the relative magnitudes of electrokinetic flow and DEP. Post shapes are contoured easily to control electric field gradients and, hence, DEP behavior. Simple simulations based on similitude of fluid flow and electric field that solve the Laplace equation to obtain fluid velocity have also been developed to predict the dielectrophoretic behavior in an array of posts. These simulations are in excellent agreement with the experimental observations and provide insight into electrokinetic behavior to enable design of dielectrophoretic concentrators and sorters.

Faceted design of channels for low-dispersion electrokinetic flows in microfluidic systems.

A novel methodology for designing microfluidic channels for low-dispersion, electrokinetic flows is presented. The technique relies on trigonometric relations that apply for ideal electrokinetic flows, allowing faceted channels to be designed using common drafting software and a hand calculator. Flows are rotated and stretched along the abrupt interface between adjacent regions having differing specific permeability--a quantity with dimensions of length that we introduce to derive the governing equations. Two-interface systems are used to eliminate hydrodynamic rotation of bands injected into channels. Regions bounded by interfaces form faceted flow "prisms" with uniform velocity fields that can be combined with other prisms to obtain a wide range of turning angles and expansion ratios. Lengths of faceted prisms can be varied arbitrarily, simplifying chip layout and allowing the ability to reduce unwanted effects such as transverse diffusion and Joule heating for a given faceted prism. Designs are demonstrated using two-dimensional numerical solutions of the Laplace equation.