Designing a sensitive and quantifiable nanocolloid assay with dielectrophoretic crossover frequencies

Dielectrophoretic nanocolloid assay is a promising technique for sensitive molecular detection and identification, as target molecule hybridization onto the probe-functionalized nanocolloids can change their surface conductance and consequently their dielectrophoretic crossover frequencies. Thus, instead of relying on surface charge density increase after hybridization, as in many capacitive and field effect transistor impedance sensing techniques, the current assay utilizes the much larger surface conductance (and dielectrophoresis crossover frequency) changes to effect sensitive detection. Herein, we present a Poisson–Boltzmann theory for surfaces with finite-size molecular probes that include the surface probe conformation, their contribution to surface charge with a proper delineation of the slip and Stern planes. The theory shows that the most sensitive nanocolloid molecular sensor corresponds to a minimum in the dielectrophoretic crossover frequency with respect to the bulk concentration of the molecular probes (oligonucleotides in our case) during nanocolloid functionalization. This minimum yields the lowest number of functionalized probes that are also fully stretched because of surface probe-probe interaction. Our theory provides the surface-bulk oligonucleotide concentration isotherm and a folding number for the surface oligonucleotide conformation from the crossover frequency, the zeta potential, and the hydrodynamic radius data.     

Solution pH change in non-uniform alternating current electric fields at frequencies above the electrode charging frequency

AC Faradaic reactions have been reported as a mechanism inducing non-ideal phenomena such as flow reversal and cell deformation in electrokinetic microfluidic systems. Prior published work described experiments in parallel electrode arrays below the electrode charging frequency (f_c), the frequency for electrical double layer charging at the electrode. However, 2D spatially non-uniform AC electric fields are required for applications such as in plane AC electroosmosis, AC electrothermal pumps, and dielectrophoresis. Many microscale experimental applications utilize AC frequencies around or above f_c. In this work, a pH sensitive fluorescein sodium salt dye was used to detect [H⁺] as an indicator of Faradaic reactions in aqueous solutions within non-uniform AC electric fields. Comparison experiments with (a) parallel (2D uniform fields) electrodes and (b) organic media were employed to deduce the electrode charging mechanism at 5 kHz (1.5f_c). Time dependency analysis illustrated that Faradaic reactions exist above the theoretically predicted electrode charging frequency. Spatial analysis showed [H⁺] varied spatially due to electric field non-uniformities and local pH changed at length scales greater than 50 μm away from the electrode surface. Thus, non-uniform AC fields yielded spatially varied pH gradients as a direct consequence of ion path length differences while uniform fields did not yield pH gradients; the latter is consistent with prior published data. Frequency dependence was examined from 5 kHz to 12 kHz at 5.5 V_pp potential, and voltage dependency was explored from 3.5 to 7.5 V_pp at 5 kHz. Results suggest that Faradaic reactions can still proceed within electrochemical systems in the absence of well-established electrical double layers. This work also illustrates that in microfluidic systems, spatial medium variations must be considered as a function of experiment time, initial medium conditions, electric signal potential, frequency, and spatial position.     

Refinement of the theory for extracting cell dielectric properties from dielectrophoresis and electrorotation experiments

A modified theory is proposed for extracting cell dielectric properties from the peak frequency measurement of electrorotation (ER) and the crossover frequency measurement of dielectrophoresis (DEP). Current theory in the literature is based on the low frequency (DC) approximations for the equivalent cell permittivity and conductivity, which are valid when the measurements are performed in a medium with conductivity less than 1 mS/m. The present theory extracts the cell properties through optimizing an expression for the medium conductivity in terms of the peak ER, or DEP crossover, frequency according to its definition using full expressions of equivalent cell permittivity and conductivity. Various levels of approximation of the theory are proposed and discussed through a scaling analysis. The present theory can extract both membrane and interior properties from the low and the high peak ER, or DEP crossover, frequencies for any medium conductivity provided the peak ER, or DEP crossover, frequency exists. It can be reduced to the linear theory for the low peak ER and DEP crossover frequencies in the literature when the medium conductivity is less than 10 mS/m. However, we can determine the membrane capacitance and conductance via the slope and intercept, respectively, of the straight line fitting of the ER peak and DEP frequency against medium conductivity data according to the linear theory only when the intercept dominates the experimental uncertainty, which occurs when the medium conductivity is less than 1 mS/m in practice.     

Modeling of dielectrophoretic transport of myoglobin molecules in microchannels

Myoglobin is one of the premature identifying cardiac markers, whose concentration increases from 90 pg∕ml or less to over 250 ng∕ml in the blood serum of human beings after minor heart attack. Separation, detection, and quantification of myoglobin play a vital role in revealing the cardiac arrest in advance, which is the challenging part of ongoing research. In the present work, one of the electrokinetic approaches, i.e., dielectrophoresis (DEP), is chosen to separate the myoglobin. A mathematical model is developed for simulating dielectrophoretic behavior of a myoglobin molecule in a microchannel to provide a theoretical basis for the above application. This model is based on the introduction of a dielectrophoretic force and a dielectric myoglobin model. A dielectric myoglobin model is developed by approximating the shape of the myoglobin molecule as sphere, oblate, and prolate spheroids. A generalized theoretical expression for the dielectrophoretic force acting on respective shapes of the molecule is derived. The microchannel considered for analysis has an array of parallel rectangular electrodes at the bottom surface. The potential and electric field distributions are calculated using Green’s theorem method and finite element method. These results also compared to the Fourier series method, closed form solutions by Morgan et al. [J. Phys. D: Appl. Phys. 34, 1553 (2001)] and Chang et al. [J. Phys. D: Appl. Phys. 36, 3073 (2003)]. It is observed that both Green’s theorem based analytical solution and finite element based numerical solution for proposed model are closely matched for electric field and square electric field gradients. The crossover frequency is obtained as 40 MHz for given properties of myoglobin and for all approximated shapes of myoglobin molecule. The effect of conductivity of medium and myoglobin on the crossover frequency is also demonstrated. Further, the effect of hydration layer on the crossover frequency of myoglobin molecules is also presented. Both positive and negative DEP effects on myoglobin molecules are obtained by switching the frequency of applied electric field. The effect of different shapes of myoglobin on DEP force is studied and no significant effect on DEP force is observed. Finally, repulsion of myoglobin molecules from the electrode plane at 1 KHz frequency and 10 V applied voltage is observed. These results provide the ability of applying DEP force for manipulating nanosized biomolecules such as myoglobin.     

ac electroosmotic pumping induced by noncontact external electrodes

Electroosmotic (EO) pumps based on dc electroosmosis is plagued by bubble generation and other electrochemical reactions at the electrodes at voltages beyond 1 V for electrolytes. These disadvantages limit their throughput and offset their portability advantage over mechanical syringe or pneumatic pumps. ac electroosmotic pumps at high frequency (>100 kHz) circumvent the bubble problem by inducing polarization and slip velocity on embedded electrodes,¹ but they require complex electrode designs to produce a net flow. We report a new high-throughput ac EO pump design based on induced-polarization on the entire channel surface instead of just on the electrodes. Like dc EO pumps, our pump electrodes are outside of the load section and form a cm-long pump unit consisting of three circular reservoirs (3 mm in diameter) connected by a 1×1 mm channel. The field-induced polarization can produce an effective Zeta potential exceeding 1 V and an ac slip velocity estimated as 1 mm∕sec or higher, both one order of magnitude higher than earlier dc and ac pumps, giving rise to a maximum throughput of 1 μl∕sec. Polarization over the entire channel surface, quadratic scaling with respect to the field and high voltage at high frequency without electrode bubble generation are the reasons why the current pump is superior to earlier dc and ac EO pumps.     

A novel actuation method of transporting droplets by using electrical charging of droplet in a dielectric fluid

We evaluate the feasibility of manipulating droplets in two dimensions by exploiting Coulombic forces acting on conductive droplets immersed in a dielectric fluid. When a droplet suspended in an immiscible fluid is located near an electrode under a dc voltage, the droplet can be charged by direct contact, by charge transfer along an electrically conducting path, or by both mechanisms. This process is called electrical charging of droplet (ECOD). This charged droplet may then be transported rapidly by exploiting Coulombic forces. We experimentally demonstrate electrical actuation of a charged droplet by applying voltage sequences. A charged droplet is two dimensionally actuated by following the direction of the electrical field signal. The droplet does not contact the surface of the microfluidic chip when it moves. This characteristic is very advantageous because treatments of the substrate surfaces of microfluidic chip become simpler. In order to test the feasibility of using ECOD in a droplet-based microreactor, electrocoalescence of two oppositely charged droplets is also studied. When two droplets approach each other due to Coulombic attraction, a liquid bridge is formed between them. We postulate that if the applied electric field is weaker than a certain critical level, the two droplets coalesce instantaneously when the charges are exchanged and redistributed through this liquid bridge.     

Direct measurements of the frequency-dependent dielectrophoresis force

Dielectrophoresis (DEP), the phenomenon of directed motion of electrically polarizable particles in a nonuniform electric field, is promising for applications in biochemical separation and filtration. For colloidal particles in suspension, the relaxation of the ionic species in the shear layer gives rise to a frequency-dependent, bidirectional DEP force in the radio frequency range. However, quantification methods of the DEP force on individual particles with the pico-Newton resolution required for the development of theories and design of device applications are lacking. We report the use of optical tweezers as a force sensor and a lock-in phase-sensitive technique for analysis of the particle motion in an amplitude modulated DEP force. The coherent detection and sensing scheme yielded not only unprecedented sensitivity for DEP force measurements, but also provided a selectivity that clearly distinguishes the pure DEP force from all the other forces in the system, including electrophoresis, electro-osmosis, heat-induced convection, and Brownian forces, all of which can hamper accurate measurements through other existing methods. Using optical tweezers-based force transducers already developed in our laboratory, we have results that quantify the frequency-dependent DEP force and the crossover frequency of individual particles with this new experimental method.     

Dynamic superconcentration at critical-point double-layer gates of conducting nanoporous granules due to asymmetric tangential fluxes

A transient 10⁶-fold concentration of double-layer counterions by a high-intensity electric field is demonstrated at the exit pole of a millimeter-sized conducting nanoporous granule that permits ion permeation. The phenomenon is attributed to a unique counterion screening dynamics that transforms half of the surface field into a converging one toward the ejecting pole. The resulting surface conduction flux then funnels a large upstream electro-osmotic convective counterion flux into the injecting hemisphere toward the zero-dimensional gate of the ejecting hemisphere to produce the superconcentration. As the concentrated counterion is ejected into the electroneutral bulk electrolyte, it attracts co-ions and produce a corresponding concentration of the co-ions. This mechanism is also shown to trap and concentrate co-ion microcolloids of micron sizes too (macroions) and hence has potential application in bead-based molecular assays.     

Label-free electronic probing of nucleic acids and proteins at the nanoscale using the nanoneedle biosensor

Detection of proteins and nucleic acids is dominantly performed using optical fluorescence based techniques, which are more costly and timely than electrical detection due to the need for expensive and bulky optical equipment and the process of fluorescent tagging. In this paper, we discuss our study of the electrical properties of nucleic acids and proteins at the nanoscale using a nanoelectronic probe we have developed, which we refer to as the Nanoneedle biosensor. The nanoneedle consists of four thin film layers: a conductive layer at the bottom acting as an electrode, an oxide layer on top, and another conductive layer on top of that, with a protective oxide above. The presence of proteins and nucleic acids near the tip results in a decrease in impedance across the sensing electrodes. There are three basic mechanisms behind the electrical response of DNA and protein molecules in solution under an applied alternating electrical field. The first change stems from modulation of the relative permittivity at the interface. The second mechanism is the formation and relaxation of the induced dipole moment. The third mechanism is the tunneling of electrons through the biomolecules. The results presented in this paper can be extended to develop low cost point-of-care diagnostic assays for the clinical setting.     

The effect of the surface functionalization and the electrolyte concentration on the electrical conductance of silica nanochannels

It is known that the conductance of nanochannels as a function of electrolyte concentration deviates from a linearly proportional relationship and approaches a value independent of the concentration as the electrolyte concentration is lowered. Most of the proposed models account for this behavior by considering a constant surface charge density and an ideal electrolyte solution. However, at low electrolyte concentrations, the ideal electrolyte approximation is no longer valid because the ions that result from the atmospheric carbon dioxide dissolution in water dominate the ionic concentration. In this paper, arrays of silica nanochannels were electrically characterized via conductance measurements. The conductance at low salt concentrations is modeled by a variable surface charge model that accounts for all ionic species in solution. This model was used to determine the variable surface charge of the bare silica nanochannels as well as of chemically modified nanochannels. The model correctly predicted the variation of the nanochannel conductance observed after silane (aminopropyldimethylethoxysilane) functionalization and single-strand DNA immobilization. Finally, pH modification of bulk KCl solutions was employed as an alternative method of changing the surface charge of silica nanochannels. Surface charge calculated from conductance measurements performed at different bulk pH values confirmed that the surface charge of the silica nanochannel walls is sensitive to the H⁺ concentration.     

Preface to Special Topic: Papers from the 13th International Conference on Surface and Colloid Science (ICSCS) and the 83rd ACS Colloid and Surface Science Symposium, Columbia University, New York, 2009

This Special Topic section is a compilation of several original contributions covering both fundamental and practical aspects of electrokinetic microfluidic phenomena that were presented during the Electrokinetics and Microfluidics sessions held at the conference.Electrokinetics is currently the mechanism of choice for the manipulation of fluids as well as colloidal and biological particles at microscale and nanoscale dimensions.¹ The popularity of electrokinetics is perhaps not so surprising as electrodes are easy to fabricate and embed into microfluidic chips, thus allowing the entire fluid and particle actuation mechanism to be completely integrated into the device. In addition, driving microfluidics with electric fields is relatively straightforward and allows for precise actuation. Nevertheless, considerable challenges remain in understanding the complex mechanisms associated with the hydrodynamics of conducting and dielectric fluids and particles under the influence of electric fields. Concomitantly, there has been an exponential increase in research and development in this field along both fundamental and applied themes in the past five years.This sustained growth in the microfluidics community of electrokinetics research has led to a sequel to the first Electrokinetic Phenomena and Microfluidics session at the 82nd ACS Colloid and Surface Science Symposium in Raleigh, NC, in 2008, and which we hope will now be a regular feature at successive ACS Colloid and Surface Science meetings. This year at the combined 2009 13th International Conference on Surface and Colloid Science (ICSCS) and the 83rd ACS Colloid and Surface Science Symposium in New York, the Electrokinetics and Microfluidics symposium proved to be extremely popular, with three keynote lectures presented by Professor Howard Stone, Professor Hsueh-Chia Chang, and Professor Thomas Healy, and 44 oral presentations. In both 2008 and 2009, Biomicrofluidics has organized a special issue to cover some of the contributions reported at these meetings.²The growing interest in using electric fields to manipulate biological entities such as cells, DNA, and even single molecules is reflected in this year’s collection of papers with dielectrophoretic (DEP) phenomena comprising the bulk of the contributions. In Ref. ³, a new theory to describe Stern layer conductance along the surface of nanocolloids is proposed, forming the basis for the derivation of a more accurate prediction of the DEP crossover frequency. This theory is then employed to determine the conformation and, hence, optimum coverage of oligonucleotides on the surface of nanocolloid functionalized molecular probes during DNA hybridization under the influence of DEP, which can be exploited for biomolecular sensing. Other fundamental DEP papers include the investigation of particle motion under DEP induced optically via a photoconductor, in which Zhu et al.⁴ characterized the frequency dependence of the motion through the synchronous velocity spectra of the particles, and a numerical study of particle trapping at the throat of converging-diverging microchannels under the influence of negative DEP using a transient arbitrary Lagrangian–Eulerian finite element method.⁵ A more practical implementation is, on the other hand, reported by Yang et al.⁶ in which the negative DEP is exploited to separate colorectal cancer cells from other cells in a microfluidic device as a demonstration of a portable cancer detection tool.Continuing along the separation theme, but with regard to DNA separation using pulsed-field gel electrophoresis aided by sparse but regularly ordered microfabricated arrays of nanoposts, is a Brownian dynamics simulation model reported by Ou et al.⁷ in which DNA channeling, which predicts that the motion of DNA is undisturbed by the presence of arrays for large spacing to DNA equilibrium size ratios and when the field lines are straight, is predicted, consistent with experimental observations. In another fundamental paper, a direct numerical simulation model is presented to predict the current-voltage relationship across conducting pores along cell membranes, which is of fundamental importance in the electroporation process.⁸We hope that you will enjoy reading the contributions in this special topic and that it encourages you to participate in future Electrokinetics and Microfluidics meetings at the ACS Colloid and Surface Science Symposia, which we definitely hope will continue on a regular basis.     

Designing ionic channels in novel carbons for electrochemical energy storage

Tremendous efforts have been dedicated to developing high-performance energy storage devices based on the micro- or nano-manipulation of novel carbon electrodes, as certain nanocarbons are perceived to have advantages such as high specific surface areas, superior electric conductivities, excellent mechanical properties and so on. In typical electrochemical electrodes, ions are intercalated/deintercalated into/from the bulk (for batteries) or adsorbed/desorbed on/from the surface (for electrochemical capacitors). Fast ionic transport, significantly determined by ionic channels in active electrodes or supporting materials, is a prerequisite for the efficient energy storage with carbons. In this report, we summarize recent design strategies for ionic channels in novel carbons and give comments on the promising features based on those carbons towards tailorable ionic channels.     

High-throughput sorting of drops in microfluidic chips using electric capacitance

We analyze a recently introduced approach for the sorting of aqueous drops with biological content immersed in oil, using a microfluidic chip that combines the functionality of electrowetting with the high throughput of two-phase flow microfluidics. In this electrostatic sorter, three co-planar electrodes covered by a thin dielectric layer are placed directly below the fluidic channel. Switching the potential of the central electrode creates an electrical guide that leads the drop to the desired outlet. The generated force, which deflects the drop, can be tuned via the voltage. The working principle is based on a contrast in conductivity between the drop and the continuous phase, which ensures successful operation even for drops of highly conductive biological media like phosphate buffered saline. Moreover, since the electric field does not penetrate the drop, its content is protected from electrical currents and Joule heating. A simple capacitive model allows quantitative prediction of the electrostatic forces exerted on drops. The maximum achievable sorting rate is determined by a competition between electrostatic and hydrodynamic forces. Sorting speeds up to 1200 per second are demonstrated for conductive drops of 160 pl in low viscosity oil.     

北京市夏季城市热岛特征及其近地层气象场分析

应用1998-2003年7、8月份的北京市10个气象台站和一个自动台站观测资料以及大气所325米气象塔的观测资料，对北京市的夏季城市热岛特征及其气象场进行了分析。结果表明：北京市夏季城市热岛现象明显，1998-2003年期间平均热岛强度达到1.64℃；强热岛主要发生在夜间，02时的平均热岛强度为2.62℃。而白天城市热岛减弱较快，14时的平均强度为0.6℃，有时甚至会出现冷岛现象；随着北京市城市化进程的不断深入，其城市热岛强度>2℃的天数也在逐年增加，最强时可达9℃；当热岛强度很强时，北京市近地层的风场会出现较明显的热岛环流（辐合），且风速较小（<2m/s）。结合大气所325米气象塔的观测资料可以发现：城市强热岛出现时，北京市的大气边界层逆温不仅强，而且逆温层顶较高。     

构造复势函数求解含裂纹载流薄板电流密度分布

构造出一种适用于求解带有圆形孔洞的载流薄板电流密度分布的势函数，利用这种势函数以及保角映射方法，求解了带有裂纹的载流薄板的电流密度分布。计算结果显示裂纹尖端附近有很强的电流集中现象，从而为研究裂纹尖端电热效应提供了一种简单有效的新方法。     

Uniform yeast cell assembly via microfluidics

This paper reports the use of microfluidic approaches for the fabrication of yeastosomes (yeast-celloidosomes) based on self-assembly of yeast cells onto liquid-solid or liquid-gas interfaces. Precise control over fluidic flows in droplet- and bubble-forming microfluidic devices allows production of monodispersed, size-selected templates. The general strategy to organize and assemble living cells is to tune electrostatic attractions between the template (gel or gas core) and the cells via surface charging. Layer-by-Layer (LbL) polyelectrolyte deposition was employed to invert or enhance charges of solid surfaces. We demonstrated the ability to produce high-quality, monolayer-shelled yeastosome structures under proper conditions when sufficient electrostatic driving forces are present. The combination of microfluidic fabrication with cell self-assembly enables a versatile platform for designing synthetic hierarchy bio-structures.     

Enabling Mg metal anodes rechargeable in conventional electrolytes by fast ionic transport interphase

Rechargeable magnesium batteries have received extensive attention as the Mg anodes possess twice the volumetric capacity of their lithium counterparts and are dendrite-free. However, Mg anodes suffer from surface passivation film in most glyme-based conventional electrolytes, leading to irreversible plating/stripping behavior of Mg. Here we report a facile and safe method to obtain a modified Mg metal anode with a Sn-based artificial layer via ion-exchange and alloying reactions. In the artificial coating layer, Mg₂Sn alloy composites offer a channel for fast ion transport and insulating MgCl₂/SnCl₂ bestows the necessary potential gradient to prevent deposition on the surface. Significant improved ion conductivity of the solid electrolyte interfaces and decreased overpotential of Mg symmetric cells in Mg(TFSI)₂/DME electrolyte are obtained. The coated Mg anodes can sustain a stable plating/stripping process over 4000 cycles at a high current density of 6 mA cm⁻². This finding provides an avenue to facilitate fast ion diffusion kinetics of Mg metal anodes in conventional electrolytes.     

Fluctuations of DNA mobility in nanofluidic entropic traps

We studied the mobility of DNA molecules driven by an electric field through a nanofluidic device containing a periodic array of deep and shallow regions termed entropic traps. The mobility of a group of DNA molecules was measured by fluorescent video microscopy. Since the depth of a shallow region is smaller than the DNA equilibrium size, DNA molecules are trapped for a characteristic time and must compress themselves to traverse the boundary between deep and shallow regions. Consistent with previous experimental results, we observed a nonlinear relationship between the mobility and electric field strength, and that longer DNA molecules have larger mobility. In repeated measurements under seemingly identical conditions, we measured fluctuations in the mobility significantly larger than expected from statistical variation. The variation was more pronounced for lower electric field strengths where the trapping time is considerable relative to the drift time. To determine the origin of these fluctuations, we investigated the dependence of the mobility on several variables: DNA concentration, ionic strength of the solvent, fluorescent dye staining ratio, electroosmotic flow, and electric field strength. The mobility fluctuations were moderately enhanced in conditions of reduced ionic strength and electroosmotic flow.     

Transmission Line Circuit Models for Dielectric Waveguide

A general procedure is outlined for obtaining single or coupled transmission line models to represent the propagation of surface wave modes in conductively unshielded dielectric waveguides. The procedure uses a homogeneous electrically or magnetically walled waveguide having the same dimensions as the dielectric of the surface waveguide, to produce a set of orthogonal eigenfunctions. These eigenfunctions are projected upon Maxwell's equations resulting in a system of transmission lines coupled together through a wave immittance, which represents the ratio of a longitudinal and a transverse field component at the dielectric-air interface. Examples are given for various modes of the dielectric slab and the dielectric rod, in particular the HE_1n modes for the latter. The transmission line models derived for these examples consist of a single trasmission line found directly by projection or reduced from a coupled transmission line model by port elimination, or of two transmission lines coupled together. All circuit models derived preserve the basic properties of surface waves (e.g. no solution below cut-off), and any of the single line models can be solved to give explicit approximate algebraic formulae for the propagation constant as a function of frequency. Numerical results show that the dispersion curves calculated from the models versus exact values are generally excellent over the entire frequency spectrum.     

AC-dielectrophoretic characterization and separation of submicron and micron particles using sidewall AgPDMS electrodes

The recent development of microfluidic “lab on a chip” devices requires the need to continuously separate submicron particles. Here, we present a PDMS microfluidic device with sidewall conducting PDMS (AgPDMS) composite electrodes capable of separating submicron particles in hydrodynamic flow. In particular, the device can service dual functions. First, the AgPDMS composite electrodes embedded in a sidewall of the device channel allow for performing AC-dielectrophoretic (DEP) characterization through direct microscopic observation of particle behavior. Characterization experiments are carried out for numerous parameters including particle size, medium conductivity, and AC field frequency to reveal important dielectrophoresis DEP information in terms of the crossover frequency and positive/negative DEP behavior under specific frequencies. Second, the device offers an advantage that sidewall AgPDMS composite electrodes can produce strong DEP effects throughout the entire channel height, and thus the robustness of the on-chip particle separation is demonstrated for continuous separation in a flowing mixture of 0.5 and 5 μm particles with 100% separation efficiency.