Magnetically controlled valve for flow manipulation in polymer microfluidic devices

A simple, external in-line valve for use in microfluidic devices constructed of polydimethylsiloxane (PDMS) is described. The actuation of the valve is based on the principle that flexible polymer walls of a liquid channel can be pressed together by the aid of a permanent magnet and a small metal bar. In the presence of a small NdFeB magnet lying below the channel of interest, the metal bar is pulled downward simultaneously pushing the thin layer of PDMS down thereby closing the channel stopping any flow of fluid. The operation of the valve is dependent on the thickness of the PDMS layer, the height of the channel, the gap between the chip and the magnet and the strength of the magnet. The microfluidic channels are completely closed to fluid flows ranging from 0.1 to 1.0 μL/min commonly used in microfluidic applications.     

A positive pressure-driven PDMS pump for fluid handling in microfluidic chips

By utilizing the high gas permeability of polydimethylsiloxane (PDMS), a simple positive pressure-driven pumping method was introduced. The pump was an aerated PDMS with a central channel in it and packing with a transfusion bottle. It could be attached to the inlet of microfluidic chip using a Teflon tube to release the air into the microfluidic system and then to create a positive pressure for driving fluid. In comparison with the degas-based PDMS pump, positive pressure-driven PDMS pump offered increased system flexibility and reduced individual device fabrication complexity due to its independence and versatility. More importantly, it offered the advantages that the PDMS pump could be wrapped in transfusion bottles to meet the readily available requirements, and it also easily assembled, which only required the user use a Teflon tube to connect a PDMS pump and a microfluidic chip. This assembly provided great freedom to meet different pumping requirements. Furthermore, this PDMS pump could offer many possible configures of pumping power by adjusting the geometries of the pump or by combining different pump modules, the adjustment of pumping capacity was investigated. To help design pumps with a suitable pumping performance, the sealing effect, pumping pressure and flow rate were also investigated. The results indicated that the performance of the positive pressure-driven PDMS pump was reliable. Finally, we demonstrated the utility of this pumping method by applying it to a PDMS-based viscometer microfluidic chip.     

A high-shear, low Reynolds number microfluidic rheometer

We present a microfluidic rheometer that uses in situ pressure sensors to measure the viscosity of liquids at low Reynolds number. Viscosity is measured in a long, straight channel using a PDMS-based microfluidic device that consists of a channel layer and a sensing membrane integrated with an array of piezoresistive pressure sensors via plasma surface treatment. The micro-pressure sensor is fabricated using conductive particles/PDMS composites. The sensing membrane maps pressure differences at various locations within the channel in order to measure the fluid shear stress in situ at a prescribed shear rate to estimate the fluid viscosity. We find that the device is capable to measure the viscosity of both Newtonian and non-Newtonian fluids for shear rates up to 10⁴ s^?1 while keeping the Reynolds number well below 1.     

On-chip CO<Subscript>2</Subscript> incubation for pocket-sized microfluidic cell culture

We have developed an on-chip CO₂ incubation system based on mass/heat transfer from aqueous solutions of bicarbonate source to cell culture media through a permeable poly(dimethylsiloxane) (PDMS) wall. Heating a carbonate-buffered bicarbonate solution successfully regulated CO₂ generation without any feedback control. Because a microfluidic cell culture chip with the incubation system does not require an external chamber or gas supply, the entire microfluidic cell culture setup becomes pocket sized. Using 5 ml of 0.8 M sodium bicarbonate with 65 mM sodium carbonate as the water jacket, the chip maintained the temperature, osmolality, and pH of 750 μl cell culture medium within physiological levels when the chip was placed on a 37°C surface. The osmolality shift and pCO₂ of the media reservoir stabilized within <5 mmol/kg and 5.0 ± 1.0% over at least 9 days. The incubation capabilities were demonstrated through microfluidic culture of COS-7 epithelial cells under an inverted microscope for 17 days.     

高聚物微流控芯片上集成化气动微阀的研制

报道了一种新型的聚甲基丙烯酸甲酯(PMMA)/聚二甲基硅氧烷(PDMS)复合芯片。该芯片采用PMMA-PDMS…PDMS-PMMA的四层构型,以在芯片上集成气动微阀。具有液路和控制通道网路的PMMA基片与PDMS弹性膜间采用不可逆封接,分别形成液路半芯片和控制半芯片,而2个半芯片则依靠PDMS膜间的粘性实现可逆封接,组成带有微阀的全芯片。这种制备方法解决了制备PMMA-PDMS-PMMA三层结构芯片的封接难题,封接过程简单可靠。其控制部分和液路部分可以单独更换,可进一步降低使用成本,尤其适合一次性应用场合。初步实验表明:该微阀具有良好的开关性能和耐用性。     

Inhibition of on-chip PCR using PDMS–glass hybrid microfluidic chips

In the course of developing a microfluidic analytical platform incorporating the polymerase chain reaction (PCR) and subsequent capillary electrophoresis (CE) analysis for a variety of bio-assays, we examined PCR inhibition through surface interactions with the chip materials. Our devices perform PCR in a three-layer chip, a glass–poly(dimethylsiloxane)–glass sandwich in which the poly(dimethylsiloxane) (PDMS, a silicone rubber) layer is used for pneumatic membrane pumping and valving of the PCR reagents. Initial on-chip PCR–CE tests of BK virus replicated in multiple uncoated chips showed variable results, usually yielding no detectable product at the target sample concentrations used. Subsequent “chip-flush” experiments, where water or reagents were flushed through a chip and subsequently incorporated in off-chip PCR, highlighted bovine serum albumin (BSA) amongst other pre-treatments, chip materials and PCR recipes as being effective in mitigating inhibition. When the BSA channel pre-coating was applied to on-chip PCR–CE experiments, a substantial improvement (10× to 40×) in signal-to-noise (S/N) of the CE product peak was conferred, and was shown with high confidence despite high S/N variability. This is the first study to quantitatively examine BSA’s ability to reduce inhibition of PCR performed on PDMS chips, and one of very few microfluidic PCR inhibition studies of any kind to use a large number of microfluidic chips (~400). The simplicity and effectiveness of our BSA coating suggest that passivating materials applied to microfluidic device channel networks may provide a viable pathway for development of bio-compatible devices with reduced complexity and cost.     

单泵控制聚焦流形态的微流控芯片的研制

研究一种单动力源、聚焦流形态可控的用于细胞排队的微流控芯片。建立了样品沟道与鞘流沟道不同长度比例、不同夹角的模型并进行了不同负压条件下聚焦流形态仿真,运用SPSS软件进行了回归分析并进行了模型优化。在芯片的微加工过程中,利用印刷电路板(PCB)制作了母板,以聚二甲基硅氧烷(PDMS)为芯片主要材料,制作了PDMS—PDMS,PDMS—玻璃及PCB—PDMS三种芯片。制作的芯片能够在单个动力源条件下控制聚焦流宽度,使不同大小的微粒及细胞呈单个排列流动。研究结果为分析不同尺寸的细胞而选择合适的样品流沟道与鞘流沟道长度、夹角等条件提供了依据,所制作的芯片也达到了廉价且实用的目的。     

A Piezoactuated Droplet-Dispensing Microfluidic Chip

A microfluidic dispensing device that is capable of generating droplets with volumes varying between 1 nL and 50 pL at an ejection frequency of up to 6 kHz is presented. In this device, a piezoactuator pushes onto an elastic membrane via piston tips; the mechanical bending of the membrane generates a pressure pulse pushing droplets out. An analytical model was developed solving bending characteristics of a plate-actuated fluidic dispensing system and used to calculate the displaced volume. The model was extended to perform stress analysis to find the optimum piston tip radius by minimizing design stresses. The optimum piston tip radius was found to be 67% of the chamber radius. The actuation force estimated using the analytical model was then used as input to a finite element model of the dispenser. A detailed numerical analysis was then performed to model the fluid flow and droplet ejection process and to find critical geometric and operating parameters. Results from both models were used together to find the best design parameters. The device contains three layers, a silicon layer sandwiched between two polydimethylsiloxane (PDMS) polymer layers. Silicon dry etching, together with PDMS soft lithography, was used to fabricate the chip. PDMS oxygen plasma bonding is used to bond the layers. Prototypes developed were successfully tested to dispense same-sized droplets repeatedly without unwanted droplets. The design allows easy expansion and simultaneous dispensing of fluids.$hfill$[2009-0099]      

Microfluidic fluorescence-activated cell sorting (μFACS) chip with integrated piezoelectric actuators for low-cost mammalian cell enrichment

A low-cost, microfluidic fluorescence-activated cell sorting (μFACS) microchip integrated with two piezoelectric lead–zirconate–titanate actuators was demonstrated for automated, high-performance mammalian cell analysis and enrichment. In this PDMS–glass device, cells were hydrodynamically focused into a single file line in the lateral direction by two sheath flows, and then interrogated with a forward scattering and confocal fluorescent detection system. The selected cells were displaced transversely into a collection channel by two piezoelectric actuators that worked in a pull–push relay manner with a minimal switching time of ~0.8 ms. High detection throughput (~2500 cells/s), high sorting rate (~1250 cells/s), and high sorting efficiency (~98%) were successfully achieved on the μFACS system. Six cell mixture samples containing 22.87% of GFP-expressing HeLa cells were consecutively analyzed and sorted on the chip, revealing a stable sorting efficiency of 97.7 ± 0.93%. In addition, cell mixtures containing 37.65 and 3.36% GFP HeLa cells were effectively enriched up to 83.82 and 78.51%, respectively, on the microchip, and an enrichment factor of 105 for the low-purity (3.36%) sample was successfully obtained. This fully enclosed, disposable microfluidic chip provides an automated platform for low-cost fluorescence-based cell detection and enrichment, and is attractive to applications where cross-contamination between runs and aerosol hazard are the primary concerns.     

Microfluidic chips for cells capture using 3-D hydrodynamic structure array

Microfluidic chips were designed and fabricated to capture cells in a relative small volume to generate the desired concentration needed for analysis. The microfluidic chips comprise three-dimensional (3-D) cell capture structures array fabricated in PDMS. The capture structure includes two layers. The first layer consists of spacers to create small gap between the upper layer and glass. The second layer is a sharp corner U-shaped compartment with sharp corners at the fore-end. And another type capture structure with Y-shaped fluidic guide has been designed. It was demonstrated that the structures can capture cells in theory, using Darcy–Weisbach equation and COMSOL Multiphysics. Then yeast cell was chosen to test the performance of the chips. The chip without fluid guides captured ~1.44 × 10⁵ cells and the capture efficiency was up to 71 %. And the chip with fluid guides captured ~5.0 × 10⁴ cells and the capture efficiency was ~25 %. The chip without fluid guides can capture more cells because the yeast cells in the chip without fluid guides are subject to larger hydrodynamic drag force.     

Stamp-and-stick room-temperature bonding technique for microdevices

Multilayer MEMS and microfluidic designs using diverse materials demand separate fabrication of device components followed by assembly to make the final device. Structural and moving components, labile bio-molecules, fluids and temperature-sensitive materials place special restrictions on the bonding processes that can be used for assembly of MEMS devices. We describe a room temperature "stamp and stick (SAS)" transfer bonding technique for silicon, glass and nitride surfaces using a UV curable adhesive. Alternatively, poly(dimethylsiloxane) (PDMS) can also be used as the adhesive; this is particularly useful for bonding PDMS devices. A thin layer of adhesive is first spun on a flat wafer. This adhesive layer is then selectively transferred to the device chip from the wafer using a stamping process. The device chip can then be aligned and bonded to other chips/wafers. This bonding process is conformal and works even on surfaces with uneven topography. This aspect is especially relevant to microfluidics, where good sealing can be difficult to obtain with channels on uneven surfaces. Burst pressure tests suggest that wafer bonds using the UV curable adhesive could withstand pressures of 700 kPa (7 atmospheres); those with PDMS could withstand 200 to 700 kPa (2-7 atmospheres) depending on the geometry and configuration of the device.     

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.     

Micro-optofluidic switch realized by 3D printing technology

This paper presents a PDMS micro-optofluidic chip that allows a laser beam to be driven directly toward a two-phase flow stream in a micro-channel while at the same time automatically, detecting the slug’s passage and stirring the laser light, without the use of any external optical devices. When the laser beam interacts with the microfluidic flow, depending on the fluid in the channel and the laser angle of incidence, a different signal level is detected. So a continuous air–water segmented flow will generate a signal that switches between two values. The device consists of a T-junction, which generates the two-phase flow, and three optical fiber insertions, which drive the input laser beam toward a selected area of the micro-channel and detects the flow stream. Three micro-channel sections of different widths were considered: 130, 250, 420 μm and the performance of the models was obtained by comparing ray-tracing simulations. The master of the device has been realized by 3D printing technology and a protocol which realizes the PDMS chip is presented. The static and dynamic characterizations, considering both single flows and two-phase flows, were carried out, and in spite of the device’s design simplicity, the sensitivity of the system to capture changes in the segmented flows and to stir the laser light in different directions was fully confirmed. The experimental tests show the possibility of obtaining satisfactory results with channel diameters in the order of 200 μm.     

Fabrication of layered polydimethylsiloxane/perfluoropolyether microfluidic devices with solvent compatibility and valve functionality

The development of multilayer soft lithography methodology has seen polydimethysiloxane (PDMS) as the preferred material for the fabrication of microfluidic devices. However, the functionality of these PDMS microfluidic chips is often limited by the poor chemical resistance of PDMS to certain solvents. Here, we propose the use of a photocurable perfluoropolyether (PFPE), specifically FOMBLIN^® MD40 PFPE, as a candidate material to provide a solvent-resistant buffer layer to make the device substantially impervious to chemically induced swelling. We first carried out a systematic study of the solvent resistance properties of FOMBLIN^® MD40 PFPE as compared with PDMS. The comparison presented here demonstrates the superiority of FOMBLIN^® MD40 PFPE over PDMS in this regard; moreover, the results permitted to categorize solvents in four different groups depending on their swelling ratio. We then present a step-by-step recipe for a novel fabrication process that uses multilayer lithography to construct a comprehensive solvent-resistant device with fluid and control channels integrated with a valve structure and also permitting easy establishment of outside connections.     

Printing 3D microfluidic chips with a 3D sugar printer

This study demonstrated how to quickly and effectively print two-dimensional (2D) and three-dimensional (3D) microfluidic chips with a low-cost 3D sugar printer. The sugar printer was modified from a desktop 3D printer by redesigning the extruder, so the melting sugar could be extruded with pneumatic driving. Sacrificial sugar lines were first printed on a base layer followed by casting polydimethylsiloxane (PDMS) onto the layer and repeating. Microchannels were then printed in the PDMS solvent, microfluidic chips dropped into hot water to dissolve the sugar lines after the PDMS was solidified, and the microfluidic chips did not need further sealing. Different types of sugar utilized for printing material were studied with results indicating that maltitol exhibited a stable flow property compared with other sugars such as caramel or sucrose. Low cost is a significant advantage of this type of sugar printer as the machine may be purchased for only approximately $800. Additionally, as demonstrated in this study, the printed 3D microfluidic chip is a useful tool utilized for cell culture, thus proving the 3D printer is a powerful tool for medical/biological research.     

A simple and cost-effective method for fabrication of integrated electronic-microfluidic devices using a laser-patterned PDMS layer

We report a simple and cost-effective method for fabricating integrated electronic-microfluidic devices with multilayer configurations. A CO₂ laser plotter was employed to directly write patterns on a transferred polydimethylsiloxane (PDMS) layer, which served as both a bonding and a working layer. The integration of electronics in microfluidic devices was achieved by an alignment bonding of top and bottom electrode-patterned substrates fabricated with conventional lithography, sputtering and lift-off techniques. Processes of the developed fabrication method were illustrated. Major issues associated with this method as PDMS surface treatment and characterization, thickness-control of the transferred PDMS layer, and laser parameters optimization were discussed, along with the examination and testing of bonding with two representative materials (glass and silicon). The capability of this method was further demonstrated by fabricating a microfluidic chip with sputter-coated electrodes on the top and bottom substrates. The device functioning as a microparticle focusing and trapping chip was experimentally verified. It is confirmed that the proposed method has many advantages, including simple and fast fabrication process, low cost, easy integration of electronics, strong bonding strength, chemical and biological compatibility, etc.     

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.     

Microfluidic separation of magnetic particles with soft magnetic microstructures

This paper demonstrates simple and cost-effective microfluidic devices for enhanced separation of magnetic particles by using soft magnetic microstructures. By injecting a mixture of iron powder and polydimethylsiloxane (PDMS) into a prefabricated channel, an iron–PDMS microstructure was fabricated next to a microfluidic channel. Placed between two external permanent magnets, the magnetized iron–PDMS microstructure induces localized and strong forces on the magnetic particles in the direction perpendicular to the fluid flow. Due to the small distance between the microstructure and the fluid channel, the localized large magnetic field gradients result a vertical force on the magnetic particles, leading to enhanced separation of the particles. Numerical simulations were developed to compute the particle trajectories and agreed well with experimental data. Systematic experiments and numerical simulation were conducted to study the effect of relevant factors on the transport of superparamagnetic particles, including the shape of iron–PDMS microstructure, mass ratio of iron–PDMS composite, width of the microfluidic channel, and average flow velocity.     

A novel three-dimensional microfluidic platform for on chip multicellular tumor spheroid formation and culture

The formation of three-dimensional (3D) multicellular cell spheroids such as microspheres and embryoid bodies has recently gained much attention as a useful cell culture technique, but few studies have investigated the suitability of glass for spheroids formation and culture. In this work, we present a novel three-dimensional microfluidic device made of poly(dimethylsiloxane) (PDMS) and glass for the easy and rapid synthesis and culture of tumor spheroid. The cell culture unit is composed of an array of microwells on the bottom of a glass plate, bigger microwells and elastomeric microchannels on the top of a PDMS plate. Cell suspension can be easily introduced into the cell culture unit and exchange with the external liquid environment by the microfluidic channels. A single tumor spheroid can be formed and cultured in each glass cell culture chamber, the surface of which was modified with poly(vinyl alcohol) to render it to be resistant to cell adhesion. As the cell culture medium could be replaced, spheroids of the human breast cancer (MCF-7) cells were cultured on the chip for 3 days, reaching the diameters of about 150 μm. Furthermore, the MCF-7 cells were successfully cultured on the chip in 2D and 3D culture modes. Results have shown that glass is well suitable for multicellular tumor spheroids culture. The established platform provides a convenient and rapid method for tumor spheroid culture, which is also adaptable for anticancer drug screening and fundamental biomedical research in cell biology.     

Solvent-free surface modification by initiated chemical vapor deposition to render plasma bonding capabilities to surfaces

A versatile solvent-free method for surface modification of various materials including both metals and polymers is described. Strong irreversible bonds were formed when substrates modified by initiated chemical vapor deposition (iCVD) of poly(1,3,5-trivinyltrimethylcyclotrisiloxane) or poly(V3D3) and exposed to an oxygen plasma were brought into contact with plasma-treated poly(dimethylsiloxane) (PDMS). The strength of these bonds was quantified by burst pressure testing microfluidic channels in the PDMS. The burst pressures of PDMS bonded to various coated substrates were in some cases comparable to that of PDMS bonded directly to PDMS. In addition, porous PTFE membrane coated with poly(V3D3) was successfully bonded to a PDMS microfluidic device and withstood pressures of over 300 mmHg. Bond strength was shown to correlate with surface roughness and quality of the bond between the coating and substrate. This work paves a methodology to fabricate microfluidic devices that include a specifically tailored membrane. Furthermore, the bonded devices exhibited hydrolytic stability; no dramatic change was observed even after immersion in water at room temperature over a period of 10 days.