Low-cost credit card-based microfluidic devices for magnetic bead immobilisation

We report a simple low-cost magnetic microfluidic device for magnetic bead separation and immobilisation. One dimensional arrays of localised high magnetic field gradients are constructed at the interfaces between regions magnetised with opposing polarities on the magnetic Fe₂O₃ composite stripes of credit cards. The localised high magnetic field gradients are employed to trap magnetic beads on the surface of the magnetic stripe, without the need for external magnetic components. A magnetic card writer was used to deterministically pattern the magnetic stripes of credit cards to define arrays of magnetic reversals. The fabrication of the device is based on PDMS to credit card bonding of simple flow channels. Experimental results demonstrate that magnetic beads can be captured with efficiencies of 85, 67 and 27 % at flow rates of 25, 50 and 100 μL min^?1, respectively. The results show that the credit card-based magnetic separator might offer an efficient, simple, low-cost alternative to traditional microfluidic magnetic separators for applications such as immunomagnetic cell separation.     

Lab-on-a-display: a new microparticle manipulation platform using a liquid crystal display (LCD)

This paper reports a new portable microfluidic platform, “lab-on-a-display,” that microparticles are manipulated by optoelectronic tweezers (OET) on a liquid crystal display (LCD). The OET has been constructed by assembling a ground layer, a liquid chamber, and a photoconductive layer. Without lens or optical alignments, the LCD image directly forms virtual electrodes on the photoconductive layer for dielectrophoretic manipulation. The lab-on-a-display was first realized by a conventional monochromatic LCD module and a light source brighter than 5,000 lux. It was successfully applied to the programmable manipulation of 45 μm polystyrene beads; more than 100 particles were transported with an optical image-driven control, following the moving edge of the image at every moment. The effects of bead size and bias voltage on the manipulation speed were also investigated. Due to the portability and compatibility for disposable applications, this new platform has potential for programmable particle manipulation or chip-based bioprocessing including cell separation and bead-based analysis.     

Controlled counter-flow motion of magnetic bead chains rolling along microchannels

We demonstrate controlled transport of superparamagnetic beads in the opposite direction of a laminar flow. A permanent magnet assembles 200 nm magnetic particles into about 200 μm long bead chains that are aligned in parallel to the magnetic field lines. Due to a magnetic field gradient, the bead chains are attracted towards the wall of a microfluidic channel. A rotation of the permanent magnet results in a rotation of the bead chains in the opposite direction to the magnet. Due to friction on the surface, the bead chains roll along the channel wall, even in counter-flow direction, up to at a maximum counter-flow velocity of 8 mm s⁻¹. Based on this approach, magnetic beads can be accurately manoeuvred within microfluidic channels. This counter-flow motion can be efficiently be used in Lab-on-a-Chip systems, e.g. for implementing washing steps in DNA purification.     

Dean flow focusing and separation of small microspheres within a narrow size range

Rapid, selective particle separation and concentration within the bacterial size range (1–3 μm) in clinical or environmental samples promises significant improvements in detection of pathogenic microorganisms in areas including diagnostics and bio-defence. It has been proposed that microfluidic Dean flow-based separation might offer simple, efficient sample clean-up: separation of larger, bioassay contaminants to prepare bioassay targets including spores, viruses and proteins. However, reports are limited to focusing spherical particles with diameters of 5 μm or above. To evaluate Dean flow separation for (1–3 μm) range samples, we employ a 20 μm width and depth, spiral microchannel. We demonstrate focusing, separation and concentration of particles with closely spaced diameters of 2.1 and 3.2 μm, significantly smaller than previously reported as separated in Dean flow devices. The smallest target, represented by 1.0 μm particles, is not focused due to the high pressures associated with focussing particles of this size; however, it is cleaned of 93 % of 3.2 μm and 87 % of 2.1 μm microparticles. Concentration increases approaching 3.5 times, close to the maximum, were obtained for 3.2 μm particles at a flow rate of 10 μl min^?1. Increasing concentration degraded separation, commencing at significantly lower concentrations than previously predicted, particularly for particles on the limit of being focused. It was demonstrated that flow separation specificity can be fine-tuned by adjustment of output pressure differentials, improving separation of closely spaced particle sizes. We conclude that Dean flow separation techniques can be effectively applied to sample clean-up within this significant microorganism size range.     

On-chip manipulation and trapping of microorganisms using a patterned magnetic pathway

We demonstrate on-chip manipulation and trapping of individual microorganisms at designated positions on a silicon surface within a microfluidic channel. Superparamagnetic beads acted as microorganism carriers. Cyanobacterium Synechocystis sp. PCC 6803 microorganisms were immobilized on amine-functionalized magnetic beads (Dynabead^® M-270 Amine) by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)–N-hydroxysulfosuccinimide coupling chemistry. The magnetic pathway was patterned lithographically such that half-disk Ni₈₀Fe₂₀ (permalloy) 5 μm elements were arranged sequentially for a length of 400 micrometers. An external rotating magnetic field of 10 mT was used to drive a translational force (maximum 70 pN) on the magnetic bead carriers proportional to the product of the field strength and its gradient along the patterned edge. Individual microorganisms immobilized on the magnetic beads (transporting objects) were directionally manipulated using a magnetic rail track, which was able to manipulate particles as a result of asymmetric forces from the curved and flat edges of the pattern on the disk. Transporting objects were then successfully trapped in a magnetic trapping station pathway. The transporting object moves two half-disk lengths in one field rotation, resulting in movement at ~24 μm s^?1 for 1 Hz rotational frequency with 5 μm pattern elements spaced with a 1 μm gap between elements.     

Computational and performance analysis of a continuous magnetophoretic bioseparation chip with alternating magnetic fields

This paper presents the modeling and optimization of a magnetophoretic bioseparation chip for isolating cells, such as circulating tumor cells from the peripheral blood. The chip consists of a continuous-flow microfluidic platform that contains locally engineered magnetic field gradients. The high-gradient magnetic field produced by the magnets is spatially non-uniform and gives rise to an attractive force on magnetic particles flowing through a fluidic channel. Simulations of the particle–fluid transport and the magnetic force are performed to predict the trajectories and capture lengths of the particles within the fluidic channel. The computational model takes into account key forces, such as the magnetic and fluidic forces and their effect on design parameters for an effective separation. The results show that the microfluidic device has the capability of separating various cells from their native environment. An experimental study is also conducted to verify and validate the simulation results. Finally, to improve the performance of the separation device, a parametric study is performed to investigate the effects of the magnetic bead size, cell size, number of beads per cell, and flow rate on the cell separation performance.     

GMI detection of magnetic-particle concentration in continuous flow

We describe a microfluidic device for the determination of the concentration of magnetic beads under continuous flow of the carrier fluid by means of the Giant Magneto-Impedance effect (GMI). The microfluidic chip is composed of a 10 μl chamber situated on top of the GMI sensing material, which is inserted in a microstrip transmission line for the impedance measurements. Two different GMI materials have been used for the experiments: an amorphous ribbon and a permalloy based multilayer, sputtered onto the same polymeric material of the microfluidic chamber. Detection tests in continuous flow have been performed using solutions containing two types of beads: one made of ferromagnetic microparticles and the other made of superparamagnetic nanoparticles. A basic calculation of the magnitude of the fringe field created by the beads assures a detectable signal on the sensors, but the experimental difficulties severely condition the measurements. The results show a promising sensitivity for both types of particles but also reveal an important number of detection problems that must be overcame before the prototype become functional.     

Hydrodynamic trap-and-release of single particles using dual-function elastomeric valves: design, fabrication, and characterization

This paper introduces a simple method for trapping and releasing single particles, such as microbeads and living cells, using dual-function elastomeric valves. Our key technique is the utilization of the elastomeric valve as a dual-function removable trap instead of a fixed trap and a separate component for releasing trapped particles, thereby enabling a simple yet effective trap-and-release of particles. We designed, fabricated, and characterized a microfluidic-based device for trapping and releasing single beads by controlling elastomeric valves driven by pneumatic pressure and a fluid flow action. The fluid flow is controlled to ensure that beads flowing in a main stream enter into a branch channel. A bead is trapped by deflected elastomeric valves positioned at the entrance of a branch channel. The trapped bead is easily released by removing the applied pressure. The trapping and releasing of single beads of 21?μm in diameter were successfully performed under an optimized pressure and flow rate ratio. Moreover, we confirmed that continuous trapping and releasing of single beads by repeatedly switching elastomeric valves enables the collection of a controllable number of beads. Our simple method can be integrated into microfluidic systems that require single or multiple particle arrays for quantitative and high-throughput assays in applications within the fields of biology and chemistry.     

Three-dimensional analysis and enhancement of continuous magnetic separation of particles in microfluidics

In continuous magnetic separation process, particles can be deflected and separated from the direction of laminar flow by means of magnetic force depending on their magnetic susceptibility and size as well as the flow rate. To analyze and control dynamic behavior of these particles flowing in microchannels, a three-dimensional numerical model was proposed and solved for obtaining the particle trajectories under the action of a gradient magnetic field and flow field. The magnetic force distribution and particle trajectories obtained were firstly verified by analytical and experimental results. Then, a detailed analysis for the enhancement of the continuous magnetic separation efficiency by optimizing the flow parameters and microchannel configurations was carried out. The results show that the separation efficiency can be greatly improved by controlling the flow rate ratio of the two fluid streams and introducing a broadened segment in the T-shaped microchannel. And it has been demonstrated to be effective through the sorting of 2-μm and 5-μm non-magnetic particles suspended in a dilute ferrofluid by a permanent magnet. The results reported could be encouraging for the design and optimization of efficient microfluidic separation systems.     

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.     

Viscosity-difference-induced asymmetric selective focusing for large stroke particle separation

We developed a new approach for particle separation by introducing viscosity difference of the sheath flows to form an asymmetric focusing of sample particle flow. This approach relies on the high-velocity gradient in the asymmetric focusing of the particle flow to generate a lift force, which plays a dominated role in the particle separation. The larger particles migrate away from the original streamline to the side of the higher relative velocity, while the smaller particles remain close to the streamline. Under high-viscosity (glycerol–water solution) and low-viscosity (PBS) sheath flows, a significant large stroke separation between the smaller (1.0 μm) and larger (9.9 μm) particles was achieved in a sample microfluidic device. We demonstrate that the flow rate and the viscosity difference of the sheath flows have an impact on the interval distance of the particle separation that affects the collected purity and on the focusing distribution of the smaller particles that affects the collected concentration. The interval distance of 293 μm (relative to the channel width: 0.281) and the focusing distribution of 112 μm (relative to the channel width: 0.107) were obtained in the 1042-μm-width separation area of the device. This separation method proposed in our work can potentially be applied to biological and medical applications due to the wide interval distance and the narrow focusing distribution of the particle separation, by easy manufacturing in a simple device.     

Modelling immunomagnetic cell capture in CFD

The separation of cells from a complex sample by immunomagnetic capture has recently obtained increased attention for microfluidic applications. Here, we present a simulation approach for immunomagnetic separation in a flow-through microfluidic environment that for the first time takes binding kinetics of beads to target cells as well as binding of multiple beads per cell into account. The approach is implemented into a computational fluid dynamics code and facilitates the tailored design of microfluidic magnetophoretic devices with an optimised separation performance. Although the specific computational model under study is constrained to a 2D geometry, appropriate parameter sets that allow for a continuous separation of cell/bead complexes from non-magnetic particles could be derived. In addition, based on magnetophoretic mobilities, a critical threshold value of beads per cell is revealed, where further binding is considerably reduced or the reaction cascade ceases.     

Maskless writing of microfluidics: Rapid prototyping of 3D microfluidics using scratch on a polymer substrate

We present a new rapid prototyping method designed for simple fabrication of 3D microfluidics using a maskless direct writing technique on polymer substrates. The entire process is enabled by a commercial cutter plotter with 10 μm resolution precision and high speed. A CAD design of top and bottom microstructures is directly written on a polymer substrate using a cutter plotter after setting up the suitable force. The smallest channel width of 20 μm was obtained with the minimum force and 100 μm from the maximum. Also the written depth increased linearly with force from 30 to 130 μm. Several 3D microfluidic devices are demonstrated using a maskless writing technique. The entire fabrication process from CAD layout to a final 3D device can be completed in 30 min outside the clean room facilities.     

Highly efficient dual-channel cytometric-detection of micron-sized particles in microfluidic device

To demonstrate the ability to efficiently count and identify suspended micron-sized particles by simultaneously detecting their fluorescence emission and light scattering in microfabricated channel, a compact configuration that used a polydimethylsiloxane (PDMS) microfabricated channel as interrogation component, hydrodynamic focusing for particle control, and a simple free-space optical setup for signal detection, was accordingly developed. Subsequently, a quantitative count of 1.013 μm diameter fluorescently labeled beads in suspension was implemented in a microfluidic device employing both fluorescence emission and light scattering at average particle throughput ranging from 83 to 416 particles/s. As a result, the detection efficiencies above 88% for both signals and correlation percentages above 97% between them were routinely achieved. In addition, it was shown that effective differentiation of 1.013 μm fluorescently labeled beads from various unlabeled beads in mixed populations of high mixing ratios had been successfully realized in this microfluidic-device-based instrumentation. Finally, the demonstrated system was used to detect fluorescein isothiocyanate (FITC) labeled nonpathogenic bacteria of Escherichia coli (E. coli) DH5α. The results showed the detection efficiencies above 89.7% for fluorescence emission and 94.5% for light scattering signals, and a correlation of 94.9% between the two signals at an average throughput of 350 cells/s have been obtained. As a comparison, the detection accuracies of the dual-channel cytometric detection of the FITC-labeled E. coli DH5α cells in the microfluidic device are approximately 84.3% and 88.8% for fluorescence emission and light scattering respectively when compared against a manual cell count using a haemocytometer as a standard.     

Enhancement of separation efficiency on continuous magnetophoresis by utilizing L/T-shaped microchannels

In this article a novel design of on-chip continuous magnetophoretic separator was proposed by utilizing the magnetic field and L-turning/T-junction effect of the flow field for high throughput applications. The motion of the magnetic bead was simulated based on Lagrangian tracking method and the separation efficiency was calculated according to the trajectories. Impact parameters including geometrical configuration, fluid velocity, magnetic flux density, magnetic bead size, and temperature on separation efficiency were discussed. The results show that both the L- and T-microchannel separators have higher separation efficiency as compared with the conventional straight-microchannel separator because of the L-turning/T-junction effect of the flow field. The separation efficiencies for L- and T-microchannel separators are 63.4 and 100%, respectively, while it is only 43.7% for straight-microchannel separator at the same conditions. Above a critical flow rate the separation efficiency drops drastically from nearly 100% to zero while this decrease is much slower for T-shaped configurations. The separation efficiency increases initially with the increase of the external magnetic flux density and keeps nearly constant at high magnetic flux density owing to saturated magnetization of the beads. It is also found that both the magnetic bead diameter and fluid temperature have great effect on the separation efficiency. The L/T-microchannel separators presented in this article are simple and efficient for magnetophoretic separation at high flow rates and thus useful for the high-efficiency on-chip enrichment of analytes with very low concentrations.     

A dielectrophoretic barrier-based microsystem for separation of microparticles

This paper presents a microfluidic system for separation of microparticles based on the use of dielectrophoretic barriers, which are constructed by aligning two layers of microelectrode structure face-to-face on the top and bottom sides of the microchannel. The energized barriers tend to prevent the particles in the flow from passing through. However, particles may penetrate the barriers if a sufficiently high flow rate is used. The flow velocity at which the particles begin to penetrate the barrier is defined as threshold velocity. Different particles are of different threshold velocities so that they can be separated. In this paper, the electrodes are configured with open ends and aligned with a certain angle to the direction of the flow. Polystyrene microbeads of different sizes (i.e., 9.6 and 16 μm in diameter) are studied in the tests. Under the experimental conditions, two particle trajectories are observed: the 9.6 μm beads penetrate the barriers and move straightly toward the fluidic outlet, while the 16 μm beads snake their way along the electrode edges at a relatively low speed. The two subpopulations of particles are separated into spatial distance of ∼10 mm within tens of seconds. The system presents a rapid and dynamic separation process within a continuous flow.     

Microparticle transfer onto pixel electrodes of 45 μm pitch on HV-CMOS chips—Simulation and experiment

Spatially selective deposition of electrically charged microparticles onto integrated circuits that generate electrical fields in programmable patterns using electrodes on their surface was previously limited to a pixel pitch of 100 μm. Now, we demonstrate spatially selective deposition onto pixels of 45 μm pitch in experiments on a test chip allowing arbitrary patterns, but being of limited size and of fixed characteristics, complemented by COMSOL simulations. Experiments on a prototype high voltage CMOS chip demonstrate the feasibility of miniaturisation in the first place, imply simulations of interest that cannot be tested experimentally and, conversely, complement the simplified simulation models by reality checks. Using COMSOL for the optimisation of the setup parameters, particles of decreasing average diameter in a number of aerosol and electrical field geometries are simulated with particular attention to minimising contamination (deposition of particles on undesirable locations). Combining these results, the average particle diameter is decreased from 10 μm to less than 3 μm and the deposition voltage is reduced from 100 V to 30 V, when using pixels with a pitch of 45 μm. Optimising these parameters allows for more than quadrupling the spot density compared to the previous chip, on which combinatorial particle deposition with minimal contamination is achieved. Peptide arrays, having been previously shown to be a major application for this method, benefit in particular, as the increase in density from 10,000 pixels/cm² to approximately 50,000 pixels/cm² promises a significant decrease in cost-per-peptide and amount of test specimens required.     

Magnetophoretic mixing for in situ immunochemical binding on magnetic beads in a microfluidic channel

Functionalized magnetic beads offer promising solutions to a host of micro-total analysis systems ranging from immunomagnetic biosensors to cell separators. Immunochemical binding of functional biochemical agents or target biomolecules serves as a key step in such applications. Here we show how magnetophoretic motion of magnetic microspheres in a microchannel is harnessed to promote in situ immunochemical binding of short DNA strands (probe oligonucleotide) on the bead surface via streptavidin–biotin bonds. Using a transverse magnetic field gradient, the particles are transported across a co-flowing analyte stream containing biotinylated probe oligonucleotides that are labeled with a Cy3-fluorophore. Quantification of the resulting biotin–streptavidin promoted binding has been achieved through fluorescence imaging of the magnetophoretically separated magnetic particles in a third stream of phosphate buffered saline. Both the experimental and numerical data indicate that for a given flow rate, the analyte binding per bead depends on the flow fraction of the co-flowing analyte stream through the microchannel, but not on the fluid viscosity. Parametric studies of the effects of fluid viscosity, analyte flow fraction, and total flow rate on the extent of binding and the overall analyte separation rate are also conducted numerically to identify favorable operating regimes of a flow-through immunomagnetic separator for biosensing, cell separation, or high-throughput applications.     

Theoretical analysis of a new, efficient microfluidic magnetic bead separator based on magnetic structures on multiple length scales

We present a theoretical analysis of a new design for microfluidic magnetic bead separation. It combines an external array of mm-sized permanent magnets with magnetization directions alternating between up and down with μm-sized soft magnetic structures integrated in the bottom of the separation channel. The concept is studied analytically for simple representative geometries and by numerical simulation of an experimentally realistic system geometry. The array of permanent magnets provides long-range magnetic forces that attract the beads to the channel bottom, while the soft magnetic elements provide strong local retaining forces that prevent captured beads from being torn loose by the fluid drag. The addition of the soft magnetic elements increases the maximum retaining force by two orders of magnitude. The design is scalable and provides an efficient and simple solution to the capture of large amounts of magnetic beads on a microsystem platform.     

Continuous-flow in-droplet magnetic particle separation in a droplet-based microfluidic platform

This paper presents a continuous-flow in-droplet magnetic particle separation in a droplet-based microfluidic device for magnetic bead-based bioassays. Two functions, electrocoalescence and magnetic particle manipulation, are performed in this device. A pair of charging metallic needles is inserted into two aqueous channels of the device. By electrostatic force, two different solutions can be merged to be mixed at a junction of droplet generation. The manipulation of magnetic particles is achieved using an externally applied magnetic field. The magnetic particles are separated by the magnetic field to one side of the droplet and extracted by splitting the droplet into two daughter droplets: one contains the majority of the magnetic particles and the other is almost devoid of magnetic particles. The applicability of the continuous-flow in-droplet magnetic particle separation is demonstrated by performing a proof-of-concept immunoassay between streptavidin-coated magnetic beads and biotin labelled with fluorescence. This approach will be useful for various biological and chemical analyses and compartmentalization of small samples.