Microfluidic chip fabrication using hot embossing and thermal bonding of COP

The application of silicon mold inserts by micro‐hot embossing molding has been explored in microfluidic chip fabrication. For the mold insert, this study employed an SU‐8 photoresist to coat the silicon wafer. Ultraviolet light was then used to expose the pattern on the SU‐8 photoresist surface. This study replicates the microstructure of the silicon mold insert by micro‐hot embossing molding. Different processing parameters (embossing temperature, embossing pressure, embossing time, and de‐molding temperature) for the cycle‐olefin polymer (COP) film of microfluidic chips are evaluated. The results showed that the most important parameter for replication of molded microfluidic chip is embossing temperature. De‐molding temperature is the most important parameter for surface roughness of the molded microfluidic chip. The microchannel is bonded with a cover by thermal bonding processing to form the sealed microfluidic chip. The bonding temperature is the most important factor in the bonding strength of the sealed microfluidic chip. Copyright © 2009 John Wiley & Sons, Ltd.     

Injection by hydrostatic pressure in conjunction with electrokinetic force on a microfluidic chip

A simple method was developed for injecting a sample on a cross-form microfluidic chip by means of hydrostatic pressure combined with electrokinetic forces. The hydrostatic pressure was generated simply by adjusting the liquid level in different reservoirs without any additional driven equipment such as a pump. Two dispensing strategies using a floating injection and a gated injection, coupled with hydrostatic pressure loading, were tested. The fluorescence observation verified the feasibility of hydrostatic pressure loading in the separation of a mixture of fluorescein sodium salt and fluorescein isothiocyanate. This method was proved to be effective in leading cells to a separation channel for single cell analysis.     

聚甲基丙烯酸甲酯微流控分析芯片的简易热压制作法

提出聚甲基丙烯酸甲酯(PMMA)微流控分析芯片的一种简易热压制作法,研究了镍基、单晶硅和玻璃3种阳模制备芯片及芯片的封合条件.采用扫描电镜(SEM)和电荷耦合检测器(CCD)对PMMA芯片的微通道及其横截面形貌进行了表征.SEM图和CCD图表明实现了热压封接.测定了PMMA芯片的伏安曲线和电渗流,其电渗流值与文献报道值基本一致.本法制作的PMMA芯片用于电泳分离Cy5荧光染料,峰高RSD为2.2%(n=11),理论塔板数7.4×10⁴m^-1.     

Poly(methyl methacrylate) CE microchips replicated from poly(dimethylsiloxane) templates for the determination of cations

A novel method for the rapid fabrication of poly(methyl methacrylate) (PMMA) microfluidic chips using poly(dimethylsiloxane) (PDMS) templates has been demonstrated. The PDMS molds were fabricated by soft lithography. The dense prepolymerized solution of methyl methacrylate containing thermal and UV initiators was allowed to polymerized between a PDMS template and a piece of a 1 mm thick commercial PMMA plate under a UV lamp. The images of microchannels on the PDMS template were precisely replicated into the synthesized PMMA substrates during the UV-initiated polymerization of the prepolymerized solution on the surface of the PMMA plate at room temperature. The polymerization could be completed within 10 min under ambient temperature. The chips were subsequently assembled by thermal bonding of the channel plate and the cover sheet. The new fabrication method obviates the need for specialized replication equipment and reduces the complexity of prototyping and manufacturing. Nearly 20 PMMA chips were replicated using a single PDMS mold. The attractive performance of the new microfluidic chips has been demonstrated by separating and detecting cations in connection with contactless conductivity detection. The fabricated PMMA microchip has also been successfully employed for the determination of potassium and sodium in environmental and biological samples.     

聚二甲基硅氧烷基质微流控芯片封接技术的研究

考察了聚二甲基硅氧烷(Polydimethylsiloxane,PDMS)预聚体与固化剂间的配比、固化温度及固化时间对PDMS芯片封接强度的影响,得出PDMS芯片封接的最佳条件基片和盖片所用PDMS预聚体与固化剂质量配比分别为10∶1与5∶1,固化温度为75℃,固化时间分别为35～50min和25～40min,封接后继续加热60min.在该条件下封接制作的微芯片历经半年50多次的分析、冲洗及抽液后未见明显损坏,足以满足一般分析任务的要求,并将芯片成功用于两种氨基酸的快速毛细管电泳分离.     

在微流控芯片上合成对甲氧基苯甲醛肟

本文用负压进样的方法, 在自制的玻璃微流控芯片中进行了对甲氧基苯甲醛和盐酸羟胺合成对甲氧基苯甲醛肟的相转移反应. 测定了不同反应时间的产率, 并与常规方法进行了比较. 讨论了相接触面积和塞流对产率的影响.     

Parylene to silicon nitride bonding for post-integration of high pressure microfluidics to CMOS devices

High pressure-rated channels allow microfluidic assays to be performed on a smaller footprint while keeping the throughput, thanks to the higher enabled flow rates, opening up perspectives for cost-effective integration of CMOS chips to microfluidic circuits. Accordingly, this study introduces an easy, low-cost and efficient method for realizing high pressure microfluidics-to-CMOS integration. First, we report a new low temperature (280 °C) Parylene-C wafer bonding technique, where O(2) plasma-treated Parylene-C bonds directly to Si(3)N(4) with an average bonding strength of 23 MPa. The technique works for silicon wafers with a nitride surface and uses a single layer of Parylene-C deposited only on one wafer, and allows microfluidic structures to be easily formed by directly bonding to the nitride passivation layer of the CMOS devices. Exploiting this technology, we demonstrated a microfluidic chip burst pressure as high as 16 MPa, while metal electrode structures on the silicon wafer remained functional after bonding.     

Hot embossing and thermal bonding of poly(methyl methacrylate) microfluidic chips using positive temperature coefficient ceramic heater

As a self-regulating heating device, positive temperature coefficient ceramic heater was employed for hot embossing and thermal bonding of poly(methyl methacrylate) microfluidic chip because it supplied constant-temperature heating without electrical control circuits. To emboss a channel plate, a piece of poly(methyl methacrylate) plate was sandwiched between a template and a microscopic glass slide on a positive temperature coefficient ceramic heater. All the assembled components were pressed between two elastic press heads of a spring-driven press while a voltage was applied to the heater for 10 min. Subsequently, the embossed poly(methyl methacrylate) plate bearing negative relief of channel networks was bonded with a piece of poly(methyl methacrylate) cover sheet to obtain a complete microchip using a positive temperature coefficient ceramic heater and a spring-driven press. High quality microfluidic chips fabricated by using the novel embossing/bonding device were successfully applied in the electrophoretic separation of three cations. Positive temperature coefficient ceramic heater indicates great promise for the low-cost production of poly(methyl methacrylate) microchips and should find wide applications in the fabrication of other thermoplastic polymer microfluidic devices.     

Fabrication of plastic microchips by hot embossing

Plastic microchips with microchannels (100 microm wide, 40 microm deep) of varying designs have been fabricated in polymethylmethacrylate by a hot embossing process using an electroform tool produced starting with silicon chip masters. Hot-embossed chips were capped with a polymethylmethacrylate top using a proprietary solvent bonding process. Holes were drilled through the top of the chip to allow access to the channels. The chips were tested with fluid and shown to fill easily. The seal between the top of the chip and the hot embossed base was effective, and there was no leakage from the channels when fluid was pumped through the microchannels. The chips were also tested with a semen sample and the plastic chip performed identically to the previous silicon-glass and glass versions of the chip. This microfabrication technique offers a viable and potentially high-volume low cost production method for fabricating transparent microchips for analytical applications.     

玻璃微-纳流控芯片的制备及在蛋白质电动富集中的应用

发展了一种以"二次刻蚀"技术制备玻璃微-纳流控芯片的新方法. 首先, 采用紫外光刻和化学湿法刻蚀技术在玻璃基片上加工微米深度的微通道; 去除剩余的光胶后, 在刻有微通道的基片上旋涂一层新的光胶; 再通过二次紫外光刻和湿法刻蚀在该基片上加工深度小于100 nm的纳通道; 最后, 采用室温键合技术, 将带有微纳结构的基片与盖片封合制成玻璃微-纳流控复合芯片. 利用本方法可以在普通化学实验室以简易的设备制得具有微-纳米复合结构的玻璃芯片. 将此玻璃微-纳流控复合芯片成功地应用于以电动离子捕集技术富集荧光素钠异硫氰酸酯(FITC)标记的人血清蛋白(HSA). 结果表明, 对于0.5 mg/mL的FITC-HSA, 30 s内富集倍率可达到200倍以上.     

Fluorescent sensor array in a microfluidic chip

Miniaturization and automation are highly important issues for the development of high-throughput processes. The area of micro total analysis systems (muTAS) is growing rapidly and the design of new schemes which are suitable for miniaturized analytical devices is of great importance. In this paper we report the immobilization of self-assembled monolayers (SAMs) with metal ion sensing properties, on the walls of glass microchannels. The parallel combinatorial synthesis of sensing SAMs in individually addressable microchannels towards the generation of optical sensor arrays and sensing chips has been developed. [figure: see text] The advantages of microfluidic devices, surface chemistry, parallel synthesis, and combinatorial approaches have been merged to integrate a fluorescent chemical sensor array in a microfluidic chip. Specifically, five different fluorescent self-assembled monolayers have been created on the internal walls of glass microchannels confined in a microfluidic chip.     

集成药物代谢微流控芯片的研制

本文研制了一种集成药物代谢微流控芯片, 此芯片可以同时完成药物代谢物的分子检测和代谢过程对药物细胞毒性的影响评价, 为进一步的药物代谢和药物相互作用研究奠定了良好的基础.     

Convenient formation of nanoparticle aggregates on microfluidic chips for highly sensitive SERS detection of biomolecules

Microfluidic chips combined with surface-enhanced Raman spectroscopy (SERS) offer an outstanding platform for rapid and high-sensitivity chemical analysis. However, it is nontrivial to conveniently form nanoparticle aggregrates (as SERS-active spots for SERS detection) in microchannels in a well-controlled manner. Here, we present a rapid, highly sensitive and label-free analytical technique for determining bovine serum albumin (BSA) on a poly(dimethylsiloxane) (PDMS) microfluidic chip using SERS. A modified PDMS pneumatic valve and nanopost arrays at the bottom of the fluidic microchannel are used for reversibly trapping gold nanoparticles to form gold aggregates, creating SERS-active spots for Raman detection. We fabricated a chip that consisted of a T-shaped fluidic channel and two modified pneumatic valves, which was suitable for fast loading of samples. Quantitative analysis of BSA is demonstrated with the measured peak intensity at 1,615 cm⁻¹ in the surface-enhanced Raman spectra. With our microfluidic chip, the detection limit of Raman can reach as low as the picomolar level, comparable to that of normal mass spectrometry.     

DNA separation with low-viscosity sieving matrix on microfabricated polycarbonate microfluidic chips

Microfluidic devices have been fabricated on polycarbonate (PC) substrates by use of a hot embossing method using a silicon master template. By adding auxiliary lines around the functional channel on the silicon master, its lifetime was significantly prolonged and the bonding strength of the PC cover plate to the microfluidic chip was greatly improved. More than 300 polycarbonate microfluidic chips have been replicated with the same silicon mold. CE separation of X-174/HaeIII DNA restriction fragments, with high resolution efficiency and good reproducibility, was achieved on these devices using the low-viscosity sieving matrix HPMC-50. Temperature was found to have a significant effect on separation efficiency.     

A low-cost, manufacturable method for fabricating capillary and optical fiber interconnects for microfluidic devices

Microfluidic chips require connections to larger macroscopic components, such as light sources, light detectors, and reagent reservoirs. In this article, we present novel methods for integrating capillaries, optical fibers, and wires with the channels of microfluidic chips. The method consists of forming planar interconnect channels in microfluidic chips and inserting capillaries, optical fibers, or wires into these channels. UV light is manually directed onto the ends of the interconnects using a microscope. UV-curable glue is then allowed to wick to the end of the capillaries, fibers, or wires, where it is cured to form rigid, liquid-tight connections. In a variant of this technique, used with light-guiding capillaries and optical fibers, the UV light is directed into the capillaries or fibers, and the UV-glue is cured by the cone of light emerging from the end of each capillary or fiber. This technique is fully self-aligned, greatly improves both the quality and the manufacturability of the interconnects, and has the potential to enable the fabrication of interconnects in a fully automated fashion. Using these methods, including a semi-automated implementation of the second technique, over 10,000 interconnects have been formed in almost 2000 microfluidic chips made of a variety of rigid materials. The resulting interconnects withstand pressures up to at least 800psi, have unswept volumes estimated to be less than 10 femtoliters, and have dead volumes defined only by the length of the capillary.     

Microchannel DNA sequencing matrices with switchable viscosities

We review the variety of thermo-responsive and shear-responsive polymer solutions with "switchable" viscosities that have been proposed for application as DNA sequencing matrices for capillary and microfluidic chip electrophoresis. Generally, highly entangled polymer solutions of high-molar mass polymers are necessary for the attainment of long DNA sequencing read lengths (> 500 bases) with short analysis times (< 3 h). However, these entangled polymer matrices create practical difficulties for microchannel electrophoresis with their extremely high viscosities, necessitating high-pressure loading into capillaries or chips. Shear-responsive (shear-thinning) polymer matrices exhibit a rapid drop in viscosity as the applied shear force is increased, but still require a high initial pressure to initiate flow of the solution into a microchannel. Polymer matrices designed to have thermo-responsive properties display either a lowered (thermo-thinning) or raised (thermo-thickening) viscosity as the temperature of the solution is elevated. These properties are generally designed into the polymers by the incorporation of moderately hydrophobic groups in some part of the polymer structure, which either phase-separate or hydrophobically aggregate at higher temperatures. In their low-viscosity states, these matrices that allow rapid loading of capillary or chip microchannels under low applied pressure. The primary goal of work in this area is to design polymer matrices that exhibit this responsive behavior and hence easy microchannel loading, without a reduction in DNA separation performance compared to conventional matrices. While good progress has been made, thermo-responsive matrices have yet to offer sequencing performance as good as nonthermo-responsive networks. The challenge remains to accomplish this goal through the innovative design of novel polymer structures.     

CE chips fabricated by injection molding and polyethylene/thermoplastic elastomer film packaging methods

This study presents a new packaging method using a polyethylene/thermoplastic elastomer (PE/TPE) film to seal an injection-molded CE chip made of either poly(methyl methacrylate) (PMMA) or polycarbonate (PC) materials. The packaging is performed at atmospheric pressure and at room temperature, which is a fast, easy, and reliable bonding method to form a sealed CE chip for chemical analysis and biomedical applications. The fabrication of PMMA and PC microfluidic channels is accomplished by using an injection-molding process, which could be mass-produced for commercial applications. In addition to microfluidic CE channels, 3-D reservoirs for storing biosamples, and CE buffers are also formed during this injection-molding process. With this approach, a commercial CE chip can be of low cost and disposable. Finally, the functionality of the mass-produced CE chip is demonstrated through its successful separation of phiX174 DNA/HaeIII markers. Experimental data show that the S/N for the CE chips using the PE/TPE film has a value of 5.34, when utilizing DNA markers with a concentration of 2 ng/microL and a CE buffer of 2% hydroxypropyl-methylcellulose (HPMC) in Tris-borate-EDTA (TBE) with 1% YO-PRO-1 fluorescent dye. Thus, the detection limit of the developed chips is improved. Lastly, the developed CE chips are used for the separation and detection of PCR products. A mixture of an amplified antibiotic gene for Streptococcus pneumoniae and phiX174 DNA/HaeIII markers was successfully separated and detected by using the proposed CE chips. Experimental data show that these DNA samples were separated within 2 min. The study proposed a promising method for the development of mass-produced CE chips.     

A surface plasmon resonance sensor on a compact disk-type microfluidic device

A surface plasmon resonance (SPR) sensor on a compact disk (CD)-type microfluidic device was developed to miniaturize the elements of a complete analytical system, pump and valves. The CD-type microfluidic device was fabricated by attaching a polydimethylsiloxane disk plate that contained microchannels and reservoirs to a flat polycarbonate disk plate that contained grating films with a thin layer of Au. The optical system of the SPR sensor and the theory for its operation are based on the principle of a grating coupled-type SPR. The sample and reagent solutions in the reservoirs on the CD-type microfluidic device were sequentially introduced into the detection chamber by centrifugal force generated by the rotation of the microfluidic device. The variation of resonance wavelength was dependent on the refractive index of the sample solution. This CD-type SPR sensor was successfully used in an immunoassay of immunoglobulin A (IgA). The anti-IgA, blocking reagent, sample and washing solution in the reservoirs were sequentially introduced into the detection chamber by changing the frequency of rotation of the microfluidic device. IgA in the sample solution was adsorbed to the anti-IgA immobilized on the Au thin layer in the detection chamber and was then detected by the SPR sensor.     

Parallel separation of multiple samples with negative pressure sample injection on a 3-D microfluidic array chip

A simple and powerful microfluidic array chip-based electrophoresis system, which is composed of a 3-D microfluidic array chip, a microvacuum pump-based negative pressure sampling device, a high-voltage supply and an LIF detector, was developed. The 3-D microfluidic array chip was fabricated with three glass plates, in which a common sample waste bus (SW(bus)) was etched in the bottom layer plate to avoid intersecting with the separation channel array. The negative pressure sampling device consists of a microvacuum air pump, a buffer vessel, a 3-way electromagnet valve, and a vacuum gauge. In the sample loading step, all the six samples and buffer solutions were drawn from their reservoirs across the injection intersections through the SW(bus) toward the common sample waste reservoir (SW(T)) by negative pressure. Only 0.5 s was required to obtain six pinched sample plugs at the channel crossings. By switching the three-way electromagnetic valve to release the vacuum in the reservoir SW(T), six sample plugs were simultaneously injected into the separation channels by EOF and electrophoretic separation was activated. Parallel separations of different analytes are presented on the 3-D array chip by using the newly developed sampling device.     

Arrays of horizontally-oriented mini-reservoirs generate steady microfluidic flows for continuous perfusion cell culture and gradient generation

This paper describes the use of arrays of horizontally-oriented reservoirs to deliver liquids through microchannels at a constant flow rate over extended periods of time (hours to days). The horizontal orientation maintains a constant hydraulic pressure drop across microfluidic channels even as the volumes of liquids within the reservoirs change over time. For a given channel-reservoir system, the magnitude of the flow velocity depends linearly on the height difference between reservoirs. The simple structure and operation mechanism make this pumping system versatile. A one-inlet-one-outlet system was used to continuously deliver media for perfusion cell culture, and an array of inlet reservoirs coupled to an outlet reservoir via microchannels was used to drive flows of multiple laminar streams. The parallel pumping scheme conveniently generated various smooth and step concentration gradients, and allowed evaluation of the effect of colchicine on myoblasts. Since the reservoir arrays are configured to be compatible with commercialized multichannel pipettors designed for 96 well plate handling, this simple pumping scheme is envisioned to be broadly useful for medium to high throughput microfluidic perfusion cell culture assays, cell migration assays, multiple laminar flow drug tests, and any other applications needing multiple microfluidic streams.