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.     

Resolving dynamics of inertial migration in straight and curved microchannels by direct cross-sectional imaging

The explosive development of inertial microfluidic systems for label-free sorting and isolation of cells demands improved understanding of the underlying physics that dictate the intriguing phenomenon of size-dependent migration in microchannels. Despite recent advances in the physics underlying inertial migration, migration dynamics in 3D is not fully understood. These investigations are hampered by the lack of easy access to the channel cross section. In this work, we report on a simple method of direct imaging of the channel cross section that is orthogonal to the flow direction using a common inverted microscope, providing vital information on the 3D cross-sectional migration dynamics. We use this approach to revisit particle migration in both straight and curved microchannels. In the rectangular channel, the high-resolution cross-sectional images unambiguously confirm the two-stage migration model proposed earlier. In the curved channel, we found two vertical equilibrium positions and elucidate the size-dependent vertical and horizontal migration dynamics. Based on these results, we propose a critical ratio of blockage ratio (β) to Dean number (De) where no net lateral migration occurs (β/De ∼ 0.01). This dimensionless number (β/De) predicts the direction of lateral migration (inward or outward) in curved and spiral channels, and thus serves as a guideline in design of such channels for particle and cell separation applications. Ultimately, the new approach to direct imaging of the channel cross section enables a wealth of previously unavailable information on the dynamics of inertial migration, which serves to improve our understanding of the underlying physics.     

Lectin-functionalized microchannels for characterizing pluripotent cells and early differentiation

Embryonic stem (ES) cells are capable of proliferating and differentiating to form cells of the three embryonic germ layers, namely, endoderm, mesoderm, and ectoderm. The utilization of human ES cell derivatives requires the ability to direct differentiation to specific lineages in defined, efficient, and scalable systems. Better markers are needed to identify early differentiation. Lectins have been reported as an attractive alternative to the common stem cell markers. They have been used to identify, characterize, and isolate various cell subpopulations on the basis of the presentation of specific carbohydrate groups on the cell surface. This article demonstrates how simple adhesion assays in lectin-coated microfluidic channels can provide key information on the interaction of lectins with ES and definitive endoderm cells and thereby track early differentiation. The microfluidic approach incorporates both binding strength and cell surface receptor density, whereas traditional flow cytometry only incorporates the latter. Both approaches are examined and shown to be complementary with the microfluidic approach providing more biologically relevant information.     

Isolation of target DNA using synergistic magnetic bead transport and electrokinetic flow

The advent and dissemination of next-generation sequencing (NGS) technologies such as Illumina''s sequencing platforms has brought forth vast reductions in the cost, time, and technical difficulties associated with DNA and RNA sequencing. Despite this trend, the workflow required to generate nucleic acid libraries for sequencing remains time-consuming and laborious. The following research proposes a method for simplifying and streamlining this process by replacing the manual washing steps of the common magnetic bead-based cleanup with a novel microfluidic method by integrating magnetic separation and electrokinetic purification (MSEP). Requiring no pumps, pipette mixing, vortexing, or centrifugation, MSEP relies on selective adsorption of target DNA onto the magnetic beads with subsequent transport of beads through a microchannel undergoing an antiparallel electroosmotic flow. The synergetic flow conditions were optimized using a simple electrohydrodynamic flow model. This work demonstrates that MSEP is as effective in eliminating adapter-dimers from the post-ligation library mix as the manual method while also greatly reducing the hands-on time and amount of pipetting required. Although MSEP has been applied specifically toward NGS library preparation at this time, it has the potential to be adapted and employed for any bead-based separation scheme, namely, solid phase extraction, sequence-specific hybridization, and immunoprecipitation on a microscale.     

Temperature-induced droplet coalescence in microchannels

This paper reports a technique for temperature-induced merging of droplets in a microchannel. The multiphase system consists of water droplet and oil as the dispersed phase and the carrying continuous phase. A resistive heater provides heating in a rectangular merging chamber. The temperature of the chamber is controlled by the voltage applied to the heater. The merging process of two neighboring droplets was investigated with different applied voltage, flow rate ratio between water and oil and total flowrate. Merging is found to be effective at high flow rate ratio, high temperature, and low total flowrate. The presented technique could be used for merging and mixing in droplet-based lab-on-a-chip platforms.     

Tuning bubbly structures in microchannels

Foams have many useful applications that arise from the structure and size distribution of the bubbles within them. Microfluidics allows for the rapid formation of uniform bubbles, where bubble size and volume fraction are functions of the input gas pressure, liquid flow rate, and device geometry. After formation, the microchannel confines the bubbles and determines the resulting foam structure. Bubbly structures can vary from a single row ("dripping"), to multiple rows ("alternating"), to densely packed bubbles ("bamboo" and dry foams). We show that each configuration arises in a distinct region of the operating space defined by bubble volume and volume fraction. We describe the boundaries between these regions using geometric arguments and show that the boundaries are functions of the channel aspect ratio. We compare these geometric arguments with foam structures observed in experiments using flow-focusing, T-junction, and co-flow designs to generate stable nitrogen bubbles in aqueous surfactant solution and stable droplets in oil containing dissolved surfactant. The outcome of this work is a set of design parameters that can be used to achieve desired foam structures as a function of device geometry and experimental control parameters.     

Vertical hydrodynamic focusing in glass microchannels

Vertical hydrodynamic focusing in microfluidic devices is investigated through simulation and through direct experimental verification using a confocal microscope and a novel form of stroboscopic imaging. Optimization for microfluidic cytometry of biological cells is examined. By combining multiple crossing junctions, it is possible to confine cells to a single analytic layer of interest. Subtractive flows are investigated as a means to move the analysis layer vertically in the channel and to correct the flatness of this layer. The simulation software (ADINA and Coventor) is shown to accurately capture the complex dependencies of the layer interfaces, which vary strongly with channel geometry and relative flow rates.     

Focused ion beam milling of microchannels in lithium niobate

We present experimental and simulation results for focused ion beam (FIB) milling of microchannels in lithium niobate in this paper. We investigate two different cuts of lithium niobate, Y- and Z-cuts, and observe that the experimental material removal rate in the FIB for both Y-cut and Z-cut samples was 0.3 μm³/nC, roughly two times greater than the material removal rate previously reported in the literature but in good agreement with the value we obtain from stopping and range of ions in matter (SRIM) simulations. Further, we investigate the FIB milling rate and resultant cross-sectional profile of microchannels at various ion beam currents and find that the milling rate decreases as a function of ion dose and correspondingly, the cross-sectional profiles change from rectangular to V-shaped. This indicates that material redeposition plays an important role at high ion dose or equivalently, high aspect ratio. We find that the experimental material removal rate decreases as a function of aspect ratio of the milled structures, in good agreement with our simulation results at low aspect ratio and in good agreement with the material removal rates previously reported in the literature at high aspect ratios. Our results show that it is indeed easier than previously assumed to fabricate nanochannels with low aspect ratio directly on lithium niobate using the FIB milling technique.     

Observation of hydrophobic-like behavior in geometrically patterned hydrophilic microchannels

We present our observation of meta-hydrophobicity, where geometrically patterned surfaces make hydrophilic microchannels exhibit hydrophobic-like behaviors. We analyze the wetting-induced energy decrease that results from the surface geometries and experimentally demonstrate how those geometries can modulate the dynamics of capillary-driven wetting and evaporation-driven drying of microfluidic systems. Our results also show that the modulated wetting dynamics can be employed to generate regulated patterns of microbubbles.     

Electroosmotic flow in vapor deposited silicon dioxide and nitride microchannels

Electroosmotic flow was studied in thin film microchannels with silicon dioxide and silicon nitride sidewalls formed using plasma-enhanced chemical vapor deposition (PECVD). A sacrificial etching process was employed for channel fabrication allowing for cross-sections with heights of 3 μm, ranging from 2 μm to 50 μm in width. Flow rates were measured for single channels and multichannel electroosmotic pump structures for pH levels ranging from 2.6 to 8.3, and zeta potentials were calculated for both silicon dioxide and silicon nitride surfaces. Flow rates as high as 0.086 μL∕min were measured for nitride multichannel pumps at applied electric fields of 300 V∕mm. The surface characteristics of PECVD nitride were analyzed and compared to more well-known oxide surfaces to determine the density of amine sites compared to silanol sites.     

Curvature-induced dielectrophoresis for continuous separation of particles by charge in spiral microchannels

The separation of particles from a heterogeneous mixture is critical in chemical and biological analyses. Many methods have been developed to separate particles in microfluidic devices. However, the majority of these separations have been limited to be size based and binary. We demonstrate herein a continuous dc electric field driven separation of carboxyl-coated and noncoated 10 μm polystyrene beads by charge in a double-spiral microchannel. This method exploits the inherent electric field gradients formed within the channel turns to manipulate particles by dielectrophoresis and is thus termed curvature-induced dielectrophoresis. The spiral microchannel is also demonstrated to continuously sort noncoated 5 μm beads, noncoated 10 μm beads, and carboxyl-coated 10 μm beads into different collecting wells by charge and size simultaneously. The observed particle separation processes in different situations are all predicted with reasonable agreements by a numerical model. This curvature-induced dielectrophoresis technique eliminates the in-channel microelectrodes and obstacles that are required in traditional electrode- and insulator-based dielectrophoresis devices. It may potentially be used to separate multiple particle targets by intrinsic properties for lab-on-a-chip applications.     

Simulations of DNA stretching by flow field in microchannels with complex geometry

Recently, we have reported the experimental results of DNA stretching by flow field in three microchannels (C. H. Lee and C. C. Hsieh, Biomicrofluidics 7(1), 014109 (2013)) designed specifically for the purpose of preconditioning DNA conformation for easier stretching. The experimental results do not only demonstrate the superiority of the new devices but also provides detailed observation of DNA behavior in complex flow field that was not available before. In this study, we use Brownian dynamics-finite element method (BD-FEM) to simulate DNA behavior in these microchannels, and compare the results against the experiments. Although the hydrodynamic interaction (HI) between DNA segments and between DNA and the device boundaries was not included in the simulations, the simulation results are in fairly good agreement with the experimental data from either the aspect of the single molecule behavior or from the aspect of ensemble averaged properties. The discrepancy between the simulation and the experimental results can be explained by the neglect of HI effect in the simulations. Considering the huge savings on the computational cost from neglecting HI, we conclude that BD-FEM can be used as an efficient and economic designing tool for developing new microfluidic device for DNA manipulation.     

Evaluation of 3D-printed molds for fabrication of non-planar microchannels

Replica obtained from micromolds patterned by simple photolithography has features with uniform heights, and attainable microchannels are thus quasi-two-dimensional. Recent progress in three-dimensional (3D) printing has enabled facile desktop fabrication of molds to replicate microchannels with varying heights. We investigated the replica obtained from four common techniques of 3D printing—fused deposition modeling, selective laser sintering, photo-polymer inkjet printing (PJ), and stereolithography (SL)—for the suitability to form microchannels in terms of the surface roughness inherent to the mechanism of 3D printing. There have been limited quantitative studies that focused on the surface roughness of a 3D-printed mold with different methods of 3D printing. We discussed that the surface roughness of the molds affected (1) transparency of the replica and (2) delamination pressure of poly(dimethylsiloxane) replica bonded to flat glass substrates. Thereafter, we quantified the accuracy of replication from 3D-printed molds by comparing the dimensions of the replicated parts to the designed dimensions and tested the ability to fabricate closely spaced microchannels. This study suggested that molds printed by PJ and SL printers were suitable for replica molding to fabricate microchannels with varying heights. The insight from this study shall be useful to fabricate 3D microchannels with controlled 3D patterns of flows guided by the geometry of the microchannels.     

Amplitude-modulated sinusoidal microchannels for observing adaptability in C. elegans locomotion

In this paper, we present a movement-based assay to observe adaptability in Caenorhabditis elegans locomotion behavior. The assay comprises a series of sinusoidal microchannels with a fixed wavelength and modulating (increasing or decreasing) amplitude. The channel width is comparable to the body diameter of the organism. Worms are allowed to enter the channel from the input port and migrate toward the output port. Within channel sections that closely match the worm's natural undulations, the worm movement is relatively quick and steady. As the channel amplitude increases or decreases along the device, the worm faces difficulty in generating the propulsive thrust, begins to slow down and eventually fails to move forward. A set of locomotion parameters (i.e., average forward velocity, number and duration of stops, range of contact angle, and cut-off region) is defined for worm locomotion in modulated sinusoidal channels and extracted from the recorded videos. The device is tested on wild-type C. elegans (N2) and two mutants (lev-8 and unc-38). We anticipate this passive, movement-based assay can be used to screen nematodes showing difference in locomotion phenotype.     

Inertial microfluidics in contraction–expansion microchannels: A review

Inertial microfluidics has brought enormous changes in the conventional cell/particle detection process and now become the main trend of sample pretreatment with outstanding throughput, low cost, and simple control method. However, inertial microfluidics in a straight microchannel is not enough to provide high efficiency and satisfying performance for cell/particle separation. A contraction–expansion microchannel is a widely used and multifunctional channel pattern involving inertial microfluidics, secondary flow, and the vortex in the chamber. The strengthened inertial microfluidics can help us to focus particles with a shorter channel length and less processing time. Both the vortex in the chamber and the secondary flow in the main channel can trap the target particles or separate particles based on their sizes more precisely. The contraction–expansion microchannels are also capable of combining with a curved, spiral, or serpentine channel to further improve the separation performance. Some recent studies have focused on the viscoelastic fluid that utilizes both elastic forces and inertial forces to separate different size particles precisely with a relatively low flow rate for the vulnerable cells. This article comprehensively reviews various contraction–expansion microchannels with Newtonian and viscoelastic fluids for particle focusing, separation, and microfluid mixing and provides particle manipulation performance data analysis for the contraction–expansion microchannel design.     

CO(2) dissolution in water using long serpentine microchannels

The evolution of carbon dioxide bubbles dissolving in water is experimentally examined using long microchannels. We study the coupling between bubble hydrodynamics and dissolution in confined geometries. The gas impregnation process in liquid produces significant flow rearrangements. Depending on the initial volumetric liquid fraction, three operating regimes are identified, namely saturating, coalescing, and dissolving. The morphological and dynamical transition from segmented to dilute bubbly flows is investigated. Tracking individual bubbles along the flow direction is used to calculate the temporal evolution of the liquid volumetric fraction and the average flow velocity near reference bubbles over long distances. This method allows us to empirically establish the functional relationship between bubble size and velocity. Finally, we examine the implication of this relationship during the coalescing flow regime, which limits the efficiency of the dissolution process.     

直流无刷电机的应用与发展前景

介绍了无刷直流电机的结构、工作原理和性能特点,分析了无刷直流电机的优越性和使用方向,展望了其发展前景。     

Characterization of microfluidic mixing and reaction in microchannels via analysis of cross-sectional patterns

For the diagnosis of biochemical reactions, the investigation of microflow behavior, and the confirmation of simulation results in microfluidics, experimentally quantitative measurements are indispensable. To characterize the mixing and reaction of fluids in microchannel devices, we propose a mixing quality index (M_qi) to quantify the cross-sectional patterns (also called mixing patterns) of fluids, captured with a confocal-fluorescence microscope (CFM). The operating parameters of the CFM for quantification were carefully tested. We analyzed mixing patterns, flow advection, and mass exchange of fluids in the devices with overlapping channels of two kinds. The mixing length of the two devices derived from the analysis of M_qi is demonstrated to be more precise than that estimated with a commonly applied method of blending dye liquors. By means of fluorescence resonance-energy transfer (FRET), we monitored the hybridization of two complementary oligonucleotides (a FRET pair) in the devices. The captured patterns reveal that hybridization is a progressive process along the downstream channel. The FRET reaction and the hybridization period were characterized through quantification of the reaction patterns. This analytical approach is a promising diagnostic tool that is applicable to the real-time analysis of biochemical and chemical reactions such as polymerase chain reaction (PCR), catalytic, or synthetic processes in microfluidic devices.