Effect of Joule heating on isoelectric focusing of proteins in a microchannel

Electric field-driven separation and purification techniques, such as isoelectric focusing (IEF) and isotachophoresis, generate heat in the system that can affect the performance of the separation process. In this study, a new mathematical model is presented for IEF that considers the temperature rise due to Joule heating. We used the model to study focusing phenomena and separation performance in a microchannel. A finite volume-based numerical technique is developed to study temperature-dependent IEF. Numerical simulation for narrow range IEF (6 < pH < 10) is performed in a straight microchannel for 100 ampholytes and two model proteins: staphylococcal nuclease and pancreatic ribonuclease. Separation results of the two proteins are obtained with and without considering the temperature rise due to Joule heating in the system for a nominal electric field of 100 V/cm. For the no Joule heating case, constant properties are used, while for the Joule heating case, temperature-dependent titration curves and thermo-physical properties are used. Our numerical results show that the temperature change due to Joule heating has a significant impact on the final focusing points of proteins, which can lower the separation performance considerably. In the absence of advection and any active cooling mechanism, the temperature increase is the highest at the mid-section of a microchannel. We also found that the maximum temperature in the system is a strong function of the ΔpK? value of the carrier ampholytes. Simulation results are also obtained for different values of applied electric fields in order to find the optimum working range considering the simulation time and buffer temperature. Moreover, the model is extended to study IEF in a straight microchip where pH is formed by supplying H⁺ and OH⁻, and the thermal analysis shows that the heat generation is negligible in ion supplied IEF.     

Asymmetry of red blood cell motions in a microchannel with a diverging and converging bifurcation

In microcirculation, red blood cells (RBCs) flowing through bifurcations may deform considerably due to combination of different phenomena that happen at the micro-scale level, such as: attraction effect, high shear, and extensional stress, all of which may influence the rheological properties and flow behavior of blood. Thus, it is important to investigate in detail the behavior of blood flow occurring at both bifurcations and confluences. In the present paper, by using a micro-PTV system, we investigated the variations of velocity profiles of two working fluids flowing through diverging and converging bifurcations, human red blood cells suspended in dextran 40 with about 14% of hematocrit level (14 Hct) and pure water seeded with fluorescent trace particles. All the measurements were performed in the center plane of rectangular microchannels using a constant flow rate of about 3.0 × 10⁻¹² m³/s. Moreover, the experimental data was compared with numerical results obtained for Newtonian incompressible fluid. The behavior of RBCs was asymmetric at the divergent and convergent side of the geometry, whereas the velocities of tracer particles suspended in pure water were symmetric and well described by numerical simulation. The formation of a red cell-depleted zone immediately downstream of the apex of the converging bifurcation was observed and its effect on velocity profiles of RBCs flow has been investigated. Conversely, a cell-depleted region was not formed around the apex of the diverging bifurcation and as a result the adhesion of RBCs to the wall surface was enhanced in this region.     

Modulation of rotation-induced lift force for cell filtration in a low aspect ratio microchannel

Cell filtration is a critical step in sample preparation in many bioapplications. Herein, we report on a simple, filter-free, microfluidic platform based on hydrodynamic inertial migration. Our approach builds on the concept of two-stage inertial migration which permits precise prediction of microparticle position within the microchannel. Our design manipulates equilibrium positions of larger microparticles by modulating rotation-induced lift force in a low aspect ratio microchannel. Here, we demonstrate filtration of microparticles with extreme efficiency (>99%). Using multiple prostate cell lines (LNCaP and human prostate epithelial tumor cells), we show filtration from spiked blood, with 3-fold concentration and >83% viability. Results of a proliferation assay show normal cell division and suggest no negative effects on intrinsic properties. Considering the planar low-aspect-ratio structure and predictable focusing, we envision promising applications and easy integration with existing lab-on-a-chip systems.     

Dielectrophoretic choking phenomenon in a converging-diverging microchannel

Experiments show that particles smaller than the throat size of converging-diverging microchannels can sometimes be trapped near the throat. This critical phenomenon is associated with the negative dc dielectrophoresis arising from nonuniform electric fields in the microchannels. A finite-element model, accounting for the particle-fluid-electric field interactions, is employed to investigate the conditions for this dielectrophoretic (DEP) choking in a converging-diverging microchannel for the first time. It is shown quantitatively that the DEP choking occurs for high nonuniformity of electric fields, high ratio of particle size to throat size, and high ratio of particle’s zeta potential to that of microchannel.     

The effect of red blood cell aggregation on velocity and cell-depleted layer characteristics of blood in a bifurcating microchannel

Red blood cell (RBC) aggregation is a multifaceted phenomenon, and whether it is generally beneficial or deleterious remains unclear. In order to better understand its effect on microvascular blood flow, the phenomenon must be studied in complex geometries, as it is strongly dependent on time, flow, and geometry. The cell-depleted layer (CDL) which forms at the walls of microvessels has been observed to be enhanced by aggregation; however, details of the characteristics of the CDL in complex regions, such as bifurcations, require further investigation. In the present study, a microchannel with a T-junction was used to analyze the influence of aggregation on the flow field and the CDL. Micro-PIV using RBCs as tracers provided high resolution cell velocity data. CDL characteristics were measured from the same data using a newly developed technique based on motion detection. Skewed and sharpened velocity profiles in the daughter branches were observed, contrary to the behavior of a continuous Newtonian fluid. RBC aggregation was observed to increase the skewness, but decrease the sharpening, of the velocity profiles in the daughter branches. The CDL width was found to be significantly greater, with a wider distribution, in the presence of aggregation and the mean width increased proportionally with the reciprocal of the fraction of flow entering the daughter branch. Aggregation also significantly increased the roughness of the interface between the CDL and the RBC core. The present results provide further insight into how RBC aggregation may affect the flow in complex geometries, which is of importance in both understanding its functions invivo, and utilizing it as a tool in microfluidic devices.     

Pressure-driven transport of particles through a converging-diverging microchannel

Pressure-driven transport of particles through a symmetric converging-diverging microchannel is studied by solving a coupled nonlinear system, which is composed of the Navier–Stokes and continuity equations using the arbitrary Lagrangian–Eulerian finite-element technique. The predicted particle translation is in good agreement with existing experimental observations. The effects of pressure gradient, particle size, channel geometry, and a particle’s initial location on the particle transport are investigated. The pressure gradient has no effect on the ratio of the translational velocity of particles through a converging-diverging channel to that in the upstream straight channel. Particles are generally accelerated in the converging region and then decelerated in the diverging region, with the maximum translational velocity at the throat. For particles with diameters close to the width of the channel throat, the usual acceleration process is divided into three stages: Acceleration, deceleration, and reacceleration instead of a monotonic acceleration. Moreover, the maximum translational velocity occurs at the end of the first acceleration stage rather than at the throat. Along the centerline of the microchannel, particles do not rotate, and the closer a particle is located near the channel wall, the higher is its rotational velocity. Analysis of the transport of two particles demonstrates the feasibility of using a converging-diverging microchannel for passive (biological and synthetic) particle separation and ordering.     

Thermal mixing of two miscible fluids in a T-shaped microchannel

In this paper, thermal mixing characteristics of two miscible fluids in a T-shaped microchannel are investigated theoretically, experimentally, and numerically. Thermal mixing processes in a T-shaped microchannel are divided into two zones, consisting of a T-junction and a mixing channel. An analytical two-dimensional model was first built to describe the heat transfer processes in the mixing channel. In the experiments, de-ionized water was employed as the working fluid. Laser induced fluorescence method was used to measure the fluid temperature field in the microchannel. Different combinations of flow rate ratios were studied to investigate the thermal mixing characteristics in the microchannel. At the T-junction, thermal diffusion is found to be dominant in this area due to the striation in the temperature contours. In the mixing channel, heat transfer processes are found to be controlled by thermal diffusion and convection. Measured temperature profiles at the T-junction and mixing channel are compared with analytical model and numerical simulation, respectively.     

Electrokinetic focusing and filtration of cells in a serpentine microchannel

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

Continuous sheath-free magnetic separation of particles in a U-shaped microchannel

Particle separation is important to many chemical and biomedical applications. Magnetic field-induced particle separation is simple, cheap, and free of fluid heating issues that accompany electric, acoustic, and optical methods. We develop herein a novel microfluidic approach to continuous sheath-free magnetic separation of particles. This approach exploits the negative or positive magnetophoretic deflection to focus and separate particles in the two branches of a U-shaped microchannel, respectively. It is applicable to both magnetic and diamagnetic particle separations, and is demonstrated through the sorting of 5 μm and 15 μm polystyrene particles suspended in a dilute ferrofluid.     

Development of assembled microchannel resonator as an alternative fabrication method of a microchannel resonator for mass sensing in flowing liquid

We propose an alternate fabrication technique of microchannel resonators based on an assembly method of three separate parts to form a microchannel resonator on a chip. The capability of the assembled microchannel resonator to detect mass is confirmed by injecting two liquids with different densities. The experimental and theoretical values of the resonator frequency shift are in agreement with each other, which confirms the consistency of the device. The noise level of the device is estimated from the Allan variance plot, so the minimum detectable mass of 230 fg after 16 s of operation is expected. By considering the time of the practical application of 1 ms, it is found that a detectable mass of around 8.51 pg is estimated, which is applicable for detecting flowing microparticles. The sub-pico to a few picogram levels of detection will be applicable for the mass analysis of flowing microparticles such as single cells and will be greatly beneficial for many fields such as chemistry, medicine, biology, and single-cell analysis.     

Viscoelastic effects on electrokinetic particle focusing in a constricted microchannel

Focusing suspended particles in a fluid into a single file is often necessary prior to continuous-flow detection, analysis, and separation. Electrokinetic particle focusing has been demonstrated in constricted microchannels by the use of the constriction-induced dielectrophoresis. However, previous studies on this subject have been limited to Newtonian fluids only. We report in this paper an experimental investigation of the viscoelastic effects on electrokinetic particle focusing in non-Newtonian polyethylene oxide solutions through a constricted microchannel. The width of the focused particle stream is found NOT to decrease with the increase in DC electric field, which is different from that in Newtonian fluids. Moreover, particle aggregations are observed at relatively high electric fields to first form inside the constriction. They can then either move forward and exit the constriction in an explosive mode or roll back to the constriction entrance for further accumulations. These unexpected phenomena are distinct from the findings in our earlier paper [Lu et al., Biomicrofluidics 8, 021802 (2014)], where particles are observed to oscillate inside the constriction and not to pass through until a chain of sufficient length is formed. They are speculated to be a consequence of the fluid viscoelasticity effects.     

Microfluidic electrical sorting of particles based on shape in a spiral microchannel

Shape is an intrinsic marker of cell cycle, an important factor for identifying a bioparticle, and also a useful indicator of cell state for disease diagnostics. Therefore, shape can be a specific marker in label-free particle and cell separation for various chemical and biological applications. We demonstrate in this work a continuous-flow electrical sorting of spherical and peanut-shaped particles of similar volumes in an asymmetric double-spiral microchannel. It exploits curvature-induced dielectrophoresis to focus particles to a tight stream in the first spiral without any sheath flow and subsequently displace them to shape-dependent flow paths in the second spiral without any external force. We also develop a numerical model to simulate and understand this shape-based particle sorting in spiral microchannels. The predicted particle trajectories agree qualitatively with the experimental observation.     

Continuous size-based separation of microparticles in a microchannel with symmetric sharp corner structures

A new microchannel with a series of symmetric sharp corner structures is reported for passive size-dependent particle separation. Micro particles of different sizes can be completely separated based on the combination of the inertial lift force and the centrifugal force induced by the sharp corner structures in the microchannel. At appropriate flow rate and Reynolds number, the centrifugal force effect on large particles, induced by the sharp corner structures, is stronger than that on small particles; hence after passing a series of symmetric sharp corner structures, large particles are focused to the center of the microchannel, while small particles are focused at two particle streams near the two side walls of the microchannel. Particles of different sizes can then be completely separated. Particle separation with this device was demonstrated using 7.32 μm and 15.5 μm micro particles. Experiments show that in comparison with the prior multi-orifice flow fractionation microchannel and multistage-multiorifice flow fractionation microchannel, this device can completely separate two-size particles with narrower particle stream band and larger separation distance between particle streams. In addition, it requires no sheath flow and complex multi-stage separation structures, avoiding the dilution of analyte sample and complex operations. The device has potentials to be used for continuous, complete particle separation in a variety of lab-on-a-chip and biomedical applications.     

Oxygen plasma treatment for reducing hydrophobicity of a sealed polydimethylsiloxane microchannel

Rapid prototyping of polydimethylsiloxane (PDMS) is often used to build microfluidic devices. However, the inherent hydrophobic nature of the material limits the use of PDMS in many applications. While different methods have been developed to transform the hydrophobic PDMS surface to a hydrophilic surface, the actual implementation proved to be time consuming due to differences in equipment and the need for characterization. This paper reports a simple and easy protocol combining a second extended oxygen plasma treatments and proper storage to produce usable hydrophilic PDMS devices. The results show that at a plasma power of 70 W, an extended treatment of over 5 min would allow the PDMS surface to remain hydrophilic for more than 6 h. Storing the treated PDMS devices in de-ionized water would allow them to maintain their hydrophilicity for weeks. Atomic force microscopy analysis shows that a longer oxygen plasma time produces a smoother surface.     

Numerical simulation on the opto-electro-kinetic patterning for rapid concentration of particles in a microchannel

This paper presents a mathematical model for laser-induced rapid electro-kinetic patterning (REP) to elucidate the mechanism for concentrating particles in a microchannel non-destructively and non-invasively. COMSOL^®(v4.2a) multiphysics software was used to examine the effect of a variety of parameters on the focusing performance of the REP. A mathematical model of the REP was developed based on the AC electrothermal flow (ACET) equations, the dielectrophoresis (DEP) equation, the energy balance equation, the Navier-Stokes equation, and the concentration-distribution equation. The medium was assumed to be a diluted solute, and different electric potentials and laser illumination were applied to the desired place. Gold (Au) electrodes were used at the top and bottom of a microchannel. For model validation, the simulation results were compared with the experimental data. The results revealed the formation of a toroidal microvortex via the ACET effect, which was generated due to laser illumination and joule-heating in the area of interest. In addition, under some conditions, such as the frequency of AC, the DEP velocity, and the particle size, the ACET force enhances and compresses resulting in the concentration of particles. The conditions of the DEP velocity and the ACET velocity are presented in detail with a comparison of the experimental results.     

An unexpected particle oscillation for electrophoresis in viscoelastic fluids through a microchannel constriction

Electrophoresis plays an important role in many applications, which, however, has so far been extensively studied in Newtonian fluids only. This work presents the first experimental investigation of particle electrophoresis in viscoelastic polyethylene oxide (PEO) solutions through a microchannel constriction under pure DC electric fields. An oscillatory particle motion is observed in the constriction region, which is distinctly different from the particle behavior in a polymer-free Newtonian fluid. This stream-wise particle oscillation continues until a sufficient number of particles form a chain to pass through the constriction completely. It is speculated that such an unexpected particle oscillating phenomenon is a consequence of the competition between electrokinetic force and viscoelastic force induced in the constriction. The electric field magnitude, particle size, and PEO concentration are all found to positively affect this viscoelasticity-related particle oscillation due to their respective influences on the two forces.     

Tunable electrochemical pH modulation in a microchannel monitored via the proton-coupled electro-oxidation of hydroquinone

Electrochemistry is a promising tool for microfluidic systems because it is relatively inexpensive, structures are simple to fabricate, and it is straight-forward to interface electronically. While most widely used in microfluidics for chemical detection or as the transduction mechanism for molecular probes, electrochemical methods can also be used to efficiently alter the chemical composition of small (typically <100 nl) microfluidic volumes in a manner that improves or enables subsequent measurements and sample processing steps. Here, solvent (H₂O) electrolysis is performed quantitatively at a microchannel Pt band electrode to increase microchannel pH. The change in microchannel pH is simultaneously tracked at a downstream electrode by monitoring changes in the i-V characteristics of the proton-coupled electro-oxidation of hydroquinone, thus providing real-time measurement of the protonated forms of hydroquinone from which the pH can be determined in a straightforward manner. Relative peak heights for protonated and deprotonated hydroquinone forms are in good agreement with expected pH changes by measured electrolysis rates, demonstrating that solvent electrolysis can be used to provide tunable, quantitative pH control within a microchannel.     

Funding for rare tumor research in China under discussion

'Throughout the world,medical research has never been equi-table.Diseases with more patients and better publicity always grasp more attention,get more funding,a...     

Entry effects of droplet in a micro confinement: Implications for deformation-based circulating tumor cell microfiltration

Deformation-based circulating tumor cell (CTC) microchips are a representative diagnostic device for early cancer detection. This type of device usually involves a process of CTC trapping in a confined microgeometry. Further understanding of the CTC flow regime, as well as the threshold passing-through pressure, is a key to the design of deformation-based CTC filtration devices. In the present numerical study, we investigate the transitional deformation and pressure signature from surface tension dominated flow to viscous shear stress dominated flow using a droplet model. Regarding whether CTC fully blocks the channel inlet, we observe two flow regimes: CTC squeezing and shearing regime. By studying the relation of CTC deformation at the exact critical pressure point for increasing inlet velocity, three different types of cell deformation are observed: (1) hemispherical front, (2) parabolic front, and (3) elongated CTC co-flowing with carrier media. Focusing on the circular channel, we observe a first increasing and then decreasing critical pressure change with increasing flow rate. By pressure analysis, the concept of optimum velocity is proposed to explain the behavior of CTC filtration and design optimization of CTC filter. Similar behavior is also observed in channels with symmetrical cross sections like square and triangular but not in rectangular channels which only results in decreasing critical pressure.