Sample preparation of chemical warfare agent simulants on a digital microfluidic (DMF) device using magnetic bead-based solid-phase extraction

In the present study, the purification and extraction of five chemical warfare agent (CWA) simulants, dimethyl methyl phosphonate, di(propylene glycol) methyl ether, methyl salicylate, triethyl phosphate, and diethyl phthalate, on a digital microfluidic (DMF) device were demonstrated. The DMF on-chip purification and extraction was performed using a magnetic bead (MB)-based diol solid-phase extraction procedure. The extracted CWA simulants were detected using both matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and gas chromatography-mass spectrometry (GC-MS). The method detection limits of the DMF-MS approach using diol-MBs were also determined. In particular, for the DMF-GC-MS, the quantitative analysis ability was validated by determining accuracy, precision, calibration curve performance, and recovery efficiency. This study clearly shows that the CWA analyses can be automated on the DMF-MS platform, thereby minimizing human involvement.     

Low-cost polymer microfluidic device for on-chip extraction of bacterial DNA

A polymer microfluidic device for on-chip extraction of bacterial DNA has been developed for molecular diagnostics. In order to manufacture a low-cost, disposable microchip, micropillar arrays of high surface-to-volume ratio (0.152 μm⁻¹) were constructed on polymethyl methacrylate (PMMA) by hot embossing with an electroformed Ni mold, and their surface was modified with SiO₂ and an organosilane compound in subsequent steps. To seal open microchannels, the organosilane layer on top plane of the micropillars was selectively removed through photocatalytic oxidation via TiO₂/UV treatment at room temperature. As a result, the underlying SiO₂ surface was exposed without deteriorating the organosilane layer coated on lateral surface of the micropillars that could serve as bacterial cell adhesion moiety. Afterwards, a plasma-treated PDMS substrate was bonded to the exposed SiO₂ surface, completing the device fabrication. To optimize manufacturing throughput and process integration, the whole fabrication process was performed at 6 inch wafer-level including polymer imprinting, organosilane coating, and bonding. Preparation of bacterial DNA was carried out with the fabricated PDMS/PMMA chip according to the following procedure: bacterial cell capture, washing, in situ lysis, and DNA elution. The polymer-based microchip presented here demonstrated similar performance to Glass/Si chip in terms of bacterial cell capture efficiency and polymerase chain reaction (PCR) compatibility.     

Extraction of genomic DNA and detection of single nucleotide polymorphism genotyping utilizing an integrated magnetic bead-based microfluidic platform

This study presents a new magnetic bead-based microfluidic platform, which integrates three major modules for rapid leukocytes purification, genomic DNA (gDNA) extraction and fast analysis of genetic gene. By utilizing microfluidic technologies and magnetic beads conjugated with CD_15/45 antibodies, leukocytes in a human whole blood sample can be first purified and concentrated, followed by extraction of gDNA utilizing surface-charge switchable, DNA-specific, magnetic beads in the lysis solution. Then, specific genes associated with genetic diseases can be amplified by an on-chip polymerase chain reaction (PCR) process automatically. The whole pretreatment process including the leukocytes purification and gDNA extraction can be performed in an automatic fashion with the incorporation of the built bio-separators consisting of microcoils array within less than 20 min. The detection of single nucleotide polymorphism (SNP) genotyping of methylenetetra-hydrofolate reductase (MTHFR) C677T region associated with an increased risk of genetic diseases was further performed to demonstrate the capability of the proposed system. The extracted gDNA can be transported into a micro PCR chamber for on-chip fast nucleic acid amplification of detection genes with minimum human intervention. Hence, the developed system may provide a powerful automated platform for pretreatment of human leukocytes, gDNA extraction and fast analysis of genetic gene.     

An aptamer-based microfluidic device for thermally controlled affinity extraction

We present a microfluidic device for specific extraction and thermally activated release of analytes using nucleic acid aptamers. The device primarily consists of a microchamber that is packed with aptamer-functionalized microbeads as a stationary phase, and integrated with a micro heater and temperature sensor. We demonstrate the device operation by performing the extraction of a metabolic analyte, adenosine monophosphate coupled with thiazole orange (TO-AMP), with high selectivity to an RNA aptamer. Controlled release of TO-AMP from the aptamer surface is then conducted at low temperatures using on-chip thermal activation. This allows isocratic analyte elution, which eliminates the use of potentially harsh reagents, and enables efficient regeneration of the aptamer surfaces when device reusability is desired.     

3D-printed microfluidic manipulation device integrated with magnetic array

This paper reported a transparent, high-precision 3D-printed microfluidic device integrated with magnet array for magnetic manipulation. A reserved groove in the device can well constrain the Halbach array or conventional alternating array. Numerical simulations and experimental data indicate that the magnetic flux density ranges from 30 to 400 mT and its gradient is about 0.2–0.4 T/m in the manipulation channel. The magnetic field parameters of Halbach array in the same location are better than the other array. Diamagnetic polystyrene beads experience a repulsive force and move away from the magnetic field source under the effect of negative magnetophoresis. It is undeniable that as the flow rate increases, the ability of Halbach array to screen particle sizes decreases. Even so, it has a good particle size discrimination at a volumetric flow rate of 1.08 mL/h, which is much larger than that of a conventional PDMS device with a single magnet. The observed particle trajectories also confirm these statements. The deflection angle is related to the magnetic field, flow rate, and particle size. This 3D-printed device integrated with Halbach array offers excellent magnetic manipulation performance.     

Porous bead-based microfluidic assay: theory and confocal microscope imaging

Functionalized, porous microbeads provide large surface area to volume ratios, allowing the capture of relatively large quantities of target molecules from complex solutions and, thus, facilitating high-sensitivity assays. While in recent years the interest in bead-based assays has been growing, only a few studies focus on mass transfer in the bead’s interior and on the binding kinetics of functionalized, porous beads. In this study, streptavidin-coated, porous agarose beads are controllably positioned within a microfluidic conduit. Biotinylated quantum dots are pumped through the conduit and used as labels to monitor target analyte binding to the beads. Confocal microscopy techniques are employed to image the concentration of the bound quantum labels as a function of position and time. Three-dimensional, finite element simulations are carried out to model the mass transfer and binding kinetics within the beads. Key thermophysical properties, such as the reduced diffusivity of the quantum dots in the agarose matrix, are determined experimentally. Experimental observations are critically compared and favorably agree with theoretical predictions. The theoretical models provide a useful tool to better our understanding of the phenomena involved and to predict how various parameters affect microbead reaction kinetics and biosensor performance.     

Rotationally controlled magneto-hydrodynamic particle handling for bead-based microfluidic assays

This work presents a novel magnetic actuation scheme for advanced particle handling on our previously introduced, centrifugal microfluidic platform for array-based analysis of individual cells and beads. The conceptually simple actuation is based on the reciprocating motion of an elastomeric membrane featuring an integrated permanent magnet and a stationary magnet aligned along the orbit of a disc-based chamber. This compression chamber is placed at the downstream end of the particle capture chamber to induce centripetally directed, hydrodynamic lift forces on particles trapped in V-shaped geometrical barriers. Towards high frequencies of rotation, the on-disc magnet ceases to follow the rapidly oscillating magnetic field, so that the magnetic actuator is disabled during the initial, sedimentation-based filling of the trap array. At reduced spin speeds, the residence time of the magnetic actuator is sufficient to displace the magnetic actuator, resulting in a flow through the V-cup array that re-distributes, and eventually fully depletes, the previously trapped beads from the array. The same magnetic deflection scheme is also demonstrated to accelerate mixing, e.g. for upstream sample preparation.     

A bead-based microfluidic system for joint detection in TORCH screening at point-of-care testing

Microsystem Technologies - A concise bead-based microfluidic system has been developed for joint detection in TORCH screening at point-of-care testing. Assisted by five functionalized polycarbonate...     

In-situ measurement of magnetic nanoparticle quantity in a microfluidic device

Microsystem Technologies - In this article we demonstrate a method for the accurate in situ determination of the quantity of the entrapped magnetic nanoparticles in the reaction chamber of a...     

A chaotic mixer for magnetic bead-based micro cell sorter

An efficient magnetic force driven mixer with simple configuration is designed, fabricated, and tested. It is designed to facilitate the mixing of magnetic beads and biomolecules in a microchannel, where mixing is unavoidably inefficient due to its low Reynolds number. With appropriate temporal variations of the force field, chaotic mixing is achieved, hence the mixing becomes effective. The mixing device consists of embedded microconductors as a magnetic field source and a microchannel that guides the streams of working fluid. It is demonstrated that a pair of integrated micro conductors provides a local magnetic field strong enough to attract nearby magnetic beads. Mixing of magnetic beads is accomplished by applying a time-dependent control signal to a row of conductors, at the Reynolds number of as low as 10/sup -2/. Two-dimensional numerical simulation has been performed to design the configuration of the channel and electrodes, which creates chaotic motion of beads. It is found that a simple two-dimensional serpentine channel geometry with the transverse electrodes is able to create the stretching and folding of material lines, which is a manifestation of chaos. The mixing pattern predicted by the simulation has been confirmed by both flow visualization and PTV (particle tracking velocimetry) in the chaotic mixer fabricated, which should greatly increase the attachment of beads onto the target biomolecules. The optimum frequency of applied control signal is searched by evaluating the Lyapunov exponent in both numerical and experimental particle tracking. It is found that the range of optimum Strouhal number is 5相似文献   

A microfluidic device for thermal particle detection

We demonstrate the use of heat to count microscopic particles. A thermal particle detector (TPD) was fabricated by combining a 500-nm-thick silicon nitride membrane containing a thin-film resistive temperature detector with a silicone elastomer microchannel. Particles with diameters of 90 and 200 μm created relative temperature changes of 0.11 and ?0.44 K, respectively, as they flowed by the sensor. A first-order lumped thermal model was developed to predict the temperature changes. Multiple particles were counted in series to demonstrate the utility of the TPD as a particle counter.     

A tissue P system and a DNA microfluidic device for solving the shortest common superstring problem

This paper describes a tissue P system for solving the Shortest Common Superstring Problem in linear time. This tissue P system is well suited for parallel and distributed implementation using a microfluidic device working with DNA strands. The approach is not based on the usual brute force generate/test technique applied in DNA computing, but it builds the space solution gradually. The possible solutions/superstrings are build step by step through the parallel distributed combination of strings using the overlapping concatenation operation. Moreover, the DNA microfluidic device solves the problem autonomously, without the need of external control or manipulation.An erratum to this article can be found at     

A magnetic microstirrer and array for microfluidic mixing

We report the development of a micromachined magnetic-bar micromixer for microscale fluid mixing in biological laboratory-on-a-chip applications. The mixer design is inspired by large scale magnetic bar mixers. A rotating magnetic field causes a single magnetic bar or an array of them to rotate rapidly within a fluid environment. A fabrication process of the magnetic bar mixer is developed. Results of fluid mixing in micro channels and chambers are investigated using experimental means and computer-aided fluid simulation.     

A microfluidic microwell device for immunomagnetic single-cell trapping

Single-cell analysis has been widely applied in various biomedical applications, such as cancer diagnostics, immune status monitoring, and drug screening. To perform an accurate and rapid cellular analysis, various magnetic-activated cell sorting techniques are available in the markets. However, large sample requirement and uneven magnetic field distribution limit its application in single-cell trapping and following analysis. To address these problems, we developed a microfluidic microwell device for immunomagnetic single-cell trapping. By adding a microwell layer between the microchannel and magnet, the magnetic field along the device becomes more uniform. Besides, magnetic beads can be retained in the array of microwell after the high-speed washing step, whereas untrapped beads would be flushed away, resulting in high single-particle trapping efficiency (62%) and purity (99.6%). To achieve large-area single-cell trapping, we introduced a “sweeping” loading protocol to further expand the single-particle trapping range. In the microwell region near to the edge of the magnet, over 3000 single magnetic beads were trapped in a 10 mm² area. Finally, we demonstrated immunomagnetic-labeled THP-1 cells can successfully be trapped at single-cell level in the microwell. The cell trapping process can be done in 10 min. We believe the platform with an accurate and efficient single-cell trapping functionality could potentially be used for various cellular analyses at the single-cell level.     

A multilayer lateral-flow microfluidic device for particle separation

Particle separation technology plays an important role in a wide range of applications as a critical sample preprocessing step for analysis. In this work, we proposed and fabricated a multilayer lateral-flow particle filtration and separation device based on polydimethylsiloxane molding and transfer bonding techniques. Particle separation capability was demonstrated by 4.5-um polystyrene bead filtration and cancer cell (SK-BR-3) retrieving. This device exhibits higher throughput compared with most active particle separation methods and is less vulnerable to membrane clogging problem. This novel multilayer particle filtration and separation device is expected to find applications in biomedical, environmental and microanalysis fields.     

DNA ligation using a disposable microfluidic device combined with a micromixer and microchannel reactor

A novel PDMS and glass-based microfluidic device consisting of a micromixer and microreactor for DNA ligation is described in this article. The new passive type planar micromixer is 10.33 mm long and composed of a straight channel integrated with nozzles and pillars, and the microreactor is composed of a serpentine channel. Mixing was enhanced by convective diffusion facilitated by the nozzles and pillars. The performance of the micromixer was analytically simulated and experimentally evaluated. The micromixer showed a good mixing efficiency of 87.7% at a 500 μL/min flow rate (Re = 66.5). DNA ligation was successfully performed using the new microfluidic device, and ligation time was shortened from 4 h to 5 min. When used for on-chip ligation, this new micromixer offers advantages of disposability and portability.