Operating regimes of a magnetic split-flow thin (SPLITT) fractionation microfluidic device for immunomagnetic separation

Magnetic bead-based immunoassays in the microfluidic format have attracted particular interest as it has several advantages over other microfluidic separation techniques. Magnetic split-flow thin fractionation (SPLITT) is a compact version of microfluidic sorting where a bidispersed or polydispersed suspension of magnetic particle–analyte conjugates can be selectively isolated into co-flowing streams of nearly monodispersed particles. Although the device offers capability of identifying and separating more than one target analytes simultaneously, its performance is sensitive to the slightest variation of the operating condition. Herein, we have numerically investigated the performance of a microscale magnetic SPLITT device. Using a coupled Eulerian–Lagrangian approach, we have evaluated the capture efficiency (CE) and separation index (SI) for each particle type collected at their designated outlet of the SPLITT device and identified the best regimes of operating parameters. While the CE figures are found to be best represented by a group variable Π, the SI values are better represented as function of the product of the group variables γ and β; the SI versus Π plots clearly separate into two basic trends: one for constant β (i.e., varying γ) and the other for constant γ (i.e., varying β). Our study prescribes the desired operating regimes of a microfluidic magnetophoretic SPLITT device in a practical immunomagnetic separation application.     

SU-8 microchannels for live cell dielectrophoresis improvements

In this work a novel highly precise SU-8 fabrication technology is employed to construct microfluidic devices for sensitive dielectrophoretic (DEP) manipulation of budding yeast cells. A benchmark microfluidic live cell sorting system is presented, and the effect of microchannel misalignment above electrode topologies on live cell DEP is discussed in detail. Simplified model of budding Saccharomyces cerevisiae yeast cell is presented and validated experimentally in fabricated microfluidic devices. A novel fabrication process enabling rapid prototyping of microfluidic devices with well-aligned integrated electrodes is presented and the process flow is described. Identical devices were produced with standard soft-lithography processes. In comparison to standard PDMS based soft-lithography, an SU-8 layer was used to construct the microchannel walls sealed by a flat sheet of PDMS to obtain the microfluidic channels. Direct bonding of PDMS to SU-8 surface was achieved by efficient wet chemical silanization combined with oxygen plasma treatment of the contact surface. The presented fabrication process significantly improved the alignment of the microstructures. While, according to the benchmark study, the standard PDMS procedure fell well outside the range required for reasonable cell sorting efficiency. In addition, PDMS delamination above electrode topologies was significantly decreased over standard soft-lithography devices. The fabrication time and costs of the proposed methodology were found to be roughly the same.

     

A microfluidic device for array patterning by perpendicular electrokinetic focusing

This paper describes a microfluidic chip in which two perpendicular laminar-flow streams can be operated to sequentially address the surface of a flow-chamber with semi-parallel sample streams. The sample streams can be controlled in position and width by the method of electrokinetic focusing. For this purpose, each of the two streams is sandwiched by two parallel sheath flow streams containing just a buffer solution. The streams are being electroosmotically pumped, allowing a simple chip design and a setup with no moving parts. Positioning of the streams was adjusted in real-time by controlling the applied voltages according to an analytical model. The perpendicular focusing gives rise to overlapping regions, which, by combinatorial (bio) chemistry, might be used for fabrication of spot arrays of immobilized proteins and other biomolecules. Since the patterning procedure is done in a closed, liquid filled flow-structure, array spots will never be exposed to air and are prevented from drying. With this device configuration, it was possible to visualize an array of 49 spots on a surface area of 1 mm². This article describes the principle, fabrication, experimental results, analytical modeling and numerical simulations of the microfluidic chip.     

Electro-osmotically controllable multi-flow microreactor

An adjustable diffusion-based microfluidic reactor is presented here, which is based on electro-osmotic guiding of reagent samples. The device consists of a laminar flow chamber with two separate reagent inlets. The position and the width of the two sample streams in the flow chamber can be controlled individually by changing the flow ratio of three parallel guiding buffer streams. Since electro-osmotic flow (EOF) is used for pumping, no external pumps or other moving parts are needed. The region where the diffusive profiles of the two sample streams overlap is used for the reactions. This overlapping region can be manipulated in a predictable way by adjusting the voltages required to generate the respective electro-osmotic flow. Reaction dynamics inside the microreactor is illustrated with a reactant pair of a fluorescent calcium tracer and a calcium chloride solution. An analytical model, which is an analogue of electrical circuits to EOF, was developed and embedded into the LabView control software, allowing real-time control of the microreactor. This paper describes the simulation, fabrication and experimental characterisation of the device.     

Development of the MHD micropump with mixing function

As μ-TAS (micro total analysis system) is developed with enhancement of MEMS technology, the growth of medical and biological research areas increases rapidly. The study on LOC (lab on a chip), which is one of the μ-TAS and has the functions of mixing and analyzing with a tiny amount of sample and reagent on one chip, is actively progressed. LOC is composed of the microfluidic components such as micromixers and micropumps. Because the flow on the microfluidic system is generally laminar, it is very difficult to mix and feed fluidic reagents efficiently. This paper presents the design and the fabrication of the MHD micropump with mixing function, in which the fluids are simultaneously mixed and pumped by coupling between Lorentz force and the moving force of an electric charge in the electric field. The advanced model was determined through the commercial CFD program after several models were modified. The numerical results have been compared with the experimental results. This study shows the proposed micropump can be made through simple fabrication procedure and has the low energy consumption.     

Precisely sized separation of multiple particles based on the dielectrophoresis gradient in the z-direction

We present a new 3D dielectrophoresis-field-flow fraction (DEP-FFF) concept to achieve precise separation of multiple particles by using AC DEP force gradient in the z-direction. The interlaced electrode array was placed at the upstream of the microchannel, which not only focused the particles into a single particle stream to be at the same starting position for further separation, but also increased the spacing between each particle by the retard effect to reduce particle–particle aggregation. An inclined electrode was also designed in back of the focusing component to continuously and precisely separate different sizes of microparticles. Different magnitudes of DEP force are induced at different positions in the z-direction of the DEP gate, which causes different penetration times and positions of particles along the inclined DEP gate. 2, 3, 4, and 6?μm polystyrene beads were precisely sized fractionation to be four particle streams based on their different threshold DEP velocities that were induced by the field gradient in the z-direction when a voltage of 6.5?V_p–p was applied at a flow rate of 0.6?μl/min. Finally, Candida albicans were also sized separated to be three populations for demonstrating the feasibility of this platform in biological applications. The results showed that a high resolution sized fractionation (only 25% size difference) of multiple particles can be achieved in this DEP-based microfluidic device by applying a single AC electrical signal.     

Methods for counting particles in microfluidic applications

Microfluidic particle counters are important tools in biomedical diagnostic applications such as flow cytometry analysis. Major methods of counting particles in microfluidic devices are reviewed in this paper. The microfluidic resistive pulse sensor advances in sensitivity over the traditional Coulter counter by improving signal amplification and noise reduction techniques. Nanopore-based methods are used for single DNA molecule analysis and the capacitance counter is useful in liquids of low electrical conductivity and in sensing the changes of cell contents. Light-scattering and light-blocking counters are better for detecting larger particles or concentrated particles. Methods of using fluorescence detection have the capability for differentiating particles of similar sizes but different types that are labeled with different fluorescent dyes. The micro particle image velocimetry method has also been used for detecting and analyzing particles in a flow field. The general limitation of microfluidic particle counters is the low throughput which needs to be improved in the future. The integration of two or more existing microfluidic particle counting techniques is required for many practical on-chip applications.     

Particle handling in straight microfluidic channels via opposing electroosmotic and pressure-driven flows

The current study presents a method for producing recirculation zones in a straight microchannel using opposing pressure-driven and electrokinetically driven flows. The interaction of these two flow streams causes flow recirculation structures, which restricts the flow passage within the microchannel and causes a nozzle-like effect, thereby increasing the separation distance between particles in the fluid stream. Theoretical and experimental investigations are performed to investigate the effects of the applied electrical field intensity on the flow recirculation size, and the nozzle-like effect, respectively. In general, the results confirm that the proposed approach provides an effective means of achieving particle acceleration and separation distance within straight microchannels, and therefore provides a viable technique for improving particle manipulation and optical detection in conventional microfluidic devices. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

uFlow: software for rational engineering of secondary flows in inertial microfluidic devices

Approaches to abstract and modularize models of fluid flow in microfluidic devices can enable predictive and rational engineering of microfluidic circuits with rapid designer feedback. The shape of co-flowing streams in the inertial flow regime has become of particular importance for new developments in high throughput microscale manufacturing, biological, and chemical research. In a process known as flow sculpting, the cross-sectional distribution of fluid elements is deformed due to the combined effects of diffusion and transverse advection, which are brought on by interaction with velocity gradients induced by sequences of pillar structures. However, the difficulty in solving the Navier–Stokes equations for complex flow-deforming geometries makes design in this space unintuitive, time-consuming, and costly. To mitigate these issues, we have efficiently embedded flow deformation operations previously relegated to high-performance computing into a free, user-friendly, and cross-platform framework called “uFlow”, to bring flow sculpting to the broader community. uFlow computes flow deformation including both advection and diffusion effects from a single pillar in 25 ms on modern consumer hardware, enabling real-time manual design and exploration of microfluidic devices, and fast visualization of 3D particles fabricated via stop flow lithography or optical transient liquid molding. Advanced numerical routines give instant access to a practically infinite set of flow transformations. We showcase uFlow’s design models, describe their implementation and usage, and validate the algorithms which allow real-time feedback with confocal imaging and cutting-edge microfluidic particle fabrication.     

Miniaturized Vision System for Microfluidic Devices

One of the biggest obstacles for lab-on-chip (LOC) devices is the miniaturization of large-scale devices and its methodologies. Miniaturization of the current microscopic technologies combined with image processing may bring significant advantages for LOC devices in the dynamic processes of sizing, positioning, flow control and cell manipulation at different time scales. Here, we propose a vision system boarded on a polydimethylsiloxane (PDMS) polymer-based chip, which can be utilized in a complex microfluidic network for continuous monitoring of mammalian egg and donor cells of sizes in the range of 10–100 μm. The developed prototype system has sufficient resolution and is accompanied with a robust detection method for cell-based microfluidic applications. To assess its performance, an image processing algorithm was applied, and the capability of the detection method was evaluated using 11- and 26-μm particles. The results show that the proposed optical system of monitoring and illumination can be effectively incorporated into PDMS structures aiming at LOC devices.     

用于细胞固定及溶液混合的微水池结构

一种用于细胞固定和溶液稀释的微流控水池结构是多层微流控芯片装置其中一层上的一个小的腔室,覆盖相对结合的另一层基底上的两条或者多条微通道.与通常的宏观流动不同,在微芯片内部的流动呈层流状态,具有确定并可控制的方向.在这个微水池结构中的流动分布对水池结构和位置的变化很敏感.通过改变水池的形状或者位置就可以实现简单的流动控制.它比传统的流动控制方法,如微阀或者压力控制,更加简单,因此具有广泛的用途.例如,通过控制液体在水池中的流向可以固定细胞到特定的位置,也可以通过不同结构层之间的快速扩散来实现快速的溶液混合.     

Experimental characterization of electrical current leakage in poly(dimethylsiloxane) microfluidic devices

Poly(dimethylsiloxane) (PDMS) is usually considered as a dielectric material and the PDMS microchannel wall can be treated as an electrically insulated boundary in an applied electric field. However, in certain layouts of microfluidic networks, electrical leakage through the PDMS microfluidic channel walls may not be negligible, which must be carefully considered in the microfluidic circuit design. In this paper, we report on the experimental characterization of the electrical leakage current through PDMS microfluidic channel walls of different configurations. Our numerical and experimental studies indicate that for tens of microns thick PDMS channel walls, electrical leakage through the PDMS wall could significantly alter the electrical field in the main channel. We further show that we can use the electrical leakage through the PDMS microfluidic channel wall to control the electrolyte flow inside the microfluidic channel and manipulate the particle motion inside the microfluidic channel. More specifically, we can trap individual particles at different locations inside the microfluidic channel by balancing the electroosmotic flow and the electrophoretic migration of the particle.     

Modeling of micro/nano particle separation in microchannels with field-flow fractionation

The cyclical electrical field-flow fractionation (CyElFFF) is a very promising separation technique for particles and biological molecules such as proteins, nucleic acids, viruses, bacteria, yeast cells, mammalian cells. But a clear understanding of the mechanism and performance prediction of this system under different operating parameters is far from completed. This research focuses on a computational investigation of particle behavior in a CyElFFF system by taking into account both electrokinetic effects and particle dynamics. The model was validated with both theory and experimental results. The effects of key parameters such as applied electric field strength and frequency, solution fluid flow rate, particle size, particle shape on separation process are addressed in a systematic way. The developed model can also be utilized in studying the behavior of spherical or non-spherical particles (such as nanowire, nanorod, and nanofiber) in other microfluidic systems.     

Chitosan microfiber fabrication using a microfluidic chip and its application to cell cultures

In this study, a poly-methyl-methacrylate (PMMA) microfluidic chip with a 45° cross-junction microchannel is fabricated using a CO₂ laser machine to generate chitosan microfibers. Chitosan solution and sodium tripolyphosphate (STPP) solution were injected into the cross-junction microchannel of the microfluidic chip. The laminar flow of the chitosan solution was generated by hydrodynamic focusing. The diameter of laminar flow, which ranged from 30 to 50 μm, was controlled by changing the ratio between chitosan solution and STPP solution flow rates in the PMMA microfluidic chip. The laminar flow of the chitosan solution was converted into chitosan microfibers with STPP solution via the cross-linking reaction; the diameter of chitosan microfibers was in the range of 50–200 μm. The chitosan microfibers were then coated with collagen for cell cultivation. The results show that the chitosan microfibers provide good growth conditions for cells. They could be used as a scaffold for cell cultures in tissue engineering applications. This novel method has advantages of ease of fabrication, simple and low-cost process.     

Miniaturized DNA amplification platform with soft-lithographically fabricated continuous-flow PCR microfluidic device on a portable temperature controller

Fabrication of 3D microfluidic devices is normally quite expensive and tedious. A strategy was established to rapidly and effectively produce multilayer 3D microfluidic chips which are made of two layers of poly(methyl methacrylate) (PMMA) sheets and three layers of double-sided pressure sensitive adhesive (PSA) tapes. The channel structures were cut in each layer by cutting plotter before assembly. The structured channels were covered by a PMMA sheet on top and a PMMA carrier which contained threads to connect with tubing. A large variety of PMMA slides and PSA tapes can easily be designed and cut with the help of a cutting plotter. The microfluidic chip was manually assembled by a simple lamination process.The complete fabrication process from device design concept to working device can be completed in minutes without the need of expensive equipment such as laser, thermal lamination, and cleanroom. This rapid frabrication method was applied for design of a 3D hydrodynamic focusing device for synthesis of gold nanoparticles (AuNPs) as proof-of-concept. The fouling of AuNPs was prevented by means of a sheath flow. Different parameters such as flow rate and concentration of reagents were controlled to achieve AuNPs of various sizes. The sheet-based fabrication method offers a possibility to create complex microfluidic devices in a rapid, cheap and easy way.

     

Micropositioning and microscopic observation of individual picoliter-sized containers within SU-8 microchannels

We describe the fabrication and application of a bioanalytical chip, made of SU-8 photoresist, comprising integrated, high aspect-ratio microfluidic channels, suitable to manipulate and investigate vesicles, cell fragments and biological cells. A central micrometer-sized aperture allows electrical particle counting and planar membrane experiments, microfluids allow (sub)micrometer-sized objects to be transported and addressed with different chemicals. Here we show how lipid vesicles are positioned with micrometer precision within the micro-channels by means of pressure and electrophoretic movement. Our approach is suited for controlling and investigating (bio)chemical synthesis and cellular signalling processes in ultrasmall individual vesicles by electro-optical techniques.     

Development of a microfabricated electrochemical-cantilever hybrid platform

The design and fabrication of a combined electrochemical-cantilever microfluidic system is described. A chip integrating cantilevers with electrodes into a microchannel is presented with the accompanying polymer flow cell. Issues such as electrical and fluid connections are addressed, electromechanical behavior in ionic solution is investigated, and two uses of the system are demonstrated. First, all cantilevers are functionalized with cysteine, to facilitate detection of Cu²⁺ ions, then one cantilever is electrochemically cleaned in situ to generate a reference cantilever for differential measurements. Two concentrations of Cu²⁺ ions are successfully measured in this way. Clean cantilevers are used to probe a solution with and without [Fe(CN)₆]^3−/4− redox couple present, demonstrating the combined voltammetric and deflection readout.     

Evaluation of passive mixing behaviors in a pillar obstruction poly(dimethylsiloxane) microfluidic mixer using fluorescence microscopy

The rapid mixing of fluids passing through a microfluidic channel is very important for various applications of microfluidic systems. It has been a great challenge to achieve highly efficient mixing in a microfluidic system because it is very difficult to generate turbulence in a submillimeter-size channel at low Reynolds numbers (Re). In this paper, we fabricated a pillar obstruction microfluidic mixer and evaluated its mixing efficiency at various flow rates. The mixing behavior of confluent streams was estimated using a fluorescence microscope. Three different sets of miscible solutions (phosphate-buffered solution, gold nanocolloids and 20% glycerol), with Rhodamine 6G aqueous solution, were used as sample laminar flows. According to our experimental results, the pillar obstruction microfluidic mixer shows an excellent mixing performance in the low Re range. Here, the mixing performance was strongly dependent on the characteristic viscosity changes of different sets of miscible solutions. The pillar obstruction microfluidic mixer designed here is expected to benefit a wide range of lab-on-a-chip applications because fabrication is very simple and the mixing efficiency is excellent at low Re.     

Development of a biological detection platform utilizing a modular microfluidic stack

The goal of this project is to build a miniaturized, user-friendly cytometry setup (Datta et al. in Microfluidic platform for education and research. COMS, Baton Rouge, 2008; Frische et al. in Development of an miniaturized flow cytometry setup for visual cell inspection and sorting. Baton Rouge, Project Report, 2008) by combining a customized, microfluidic device with visual microscope inspection to detect and extract specific cells from a continuous sample flow. We developed a cytological tool, based on the Coulter particle counter principle, using a microelectrode array patterned on a borosilicate glass chip as electrical detection set-up which is fully embedded into a polymeric multi-layer microfluidic stack. The detection takes place between pairs of coplanar Cr/Au microelectrodes by sensing an impedance change caused by particles continuously carried within a microfluidic channel across the detection area under laminar flow conditions. A wide frequency range available for counting provides information on cell size, membrane capacitance, cytoplasm conductivity and is potentially of interest for in-depth cell diagnostic e.g. to detect damaged or cancerous cells and select them for extraction and further in-depth analysis.     

Experimental and numerical investigation on particle–particle interaction of multi-particle separation in an alternating and repetitive microchannel

Inertial focusing plays a major role in size-based cell separation or enrichment for microfluidic applications in many medical areas such as diagnostics and treatment. These applications often deal with suspensions of different particles which cause interactions between particles with different diameters such as particle–particle collision. In this study, particle–particle interaction in a laminar flow through a low aspect ratio alternating and repetitive microchannel is investigated both numerically and experimentally. It is revealed that particle–particle collision affects high quality particle focusing. computational fluid dynamics simulations are conducted for demonstrating the effect of the flow field in the transverse cross-section on the focusing quality and position. The experiments and simulations both revealed that if the flow is seeded with a mixture of particles of 3.3 and 9.9 µm diameters, the quality of focusing intensity is degenerated compared to the focusing features obtained by particles with a diameter of 9.9 µm solely. The results clearly show that particle–particle collision between the 3.3 and 9.9 µm particles has a negative effect on particle focusing behavior of the 9.9 µm particles.