Nanoliter dispensing method by degassed poly(dimethylsiloxane) microchannels and its application in protein crystallization

This paper describes a method of dispensing a nanoliter volume of liquid into arrays of microwells through degassed poly(dimethylsiloxane) (PDMS) microchannels. In this method, the PDMS microchannels were reversibly bound to arrays of microwells. The PDMS elastomer was predegassed and served as an internal vacuum pumping source. Various aqueous solutions were infused into arrays of microwells through the reversibly sealed PDMS microchannels. Microwells fabricated in PDMS, poly(methyl methacrylate) (PMMA), and glass were all compatible with this dispensing method. By removing the PDMS microchannels, arrays of droplets confined in the microwells were obtained. Multiplex reaction and screening at the nanoliter scale were carried out by binding two such arrays of microwells to form microchambers. We applied this method to screening the crystallization conditions of four known proteins. Long-term incubation of over 2 months was achieved by employing glass microwells. An unknown protein was then crystallized using the screening method in microwells. The crystals with sufficient size were harvested from the reversibly bound microwells. X-ray diffraction with a resolution of 3.1 Angstrom was obtained.     

Facile patterning of reduced graphene oxide film into microelectrode array for highly sensitive sensing

In this study, we develop a new technique to fabricate a reduced graphene oxide (rGO)-based microelectrode array (MEA) with low-cost soft lithography. To prepare patterned rGO, a polydimethylsiloxane (PDMS) mold with an array of microwells on its surface is fabricated using soft lithography, and GO is assembled on an indium tin oxide (ITO) electrode with a layer-by-layer method. The rGO pattern is formed by closely contacting the assembled GO film onto the ITO electrode with the PDMS mold filled with hydrazine solution in the microwells to selectively reduce the localized GO into the rGO. The MEA with patterned rGO as the microelectrode is characterized with Kelvin probe force microscopy (KFM), atomic force microscopy (AFM), and cyclic voltammetry (CV) with ferricyanide in aqueous solution as the redox probe. The KFM and AFM results demonstrate that each rGO pattern prepared under the present conditions is 3 μm in diameter, which is close to that of the PDMS mold we use. The CV results show that the rGO patterned onto the ITO exhibits a sigmoid-shaped voltammogram up to 200 mVs(-1) with a microampere level current response, suggesting that the rGO-based electrode fabricated with soft lithography behalves like a MEA. To demonstrate the potential electroanalytical application of the rGO-based MEA, prussian blue (PB) is electrodeposited onto the rGO-based MEA to form the PB/rGO-based MEA. Electrochemical studies on the formed PB/rGO-based MEA reveal that MEA shows a lower detection limit and a larger current density for the detection of H(2)O(2), as compared with the macroscopic rGO electrode. The method demonstrated here provides a simple and low-cost strategy for the fabrication of graphene-based MEA that are useful for electroanalytical applications.     

Fabrication of microfluidic devices containing patterned microwell arrays

A rapid fabrication and prototyping technique to incorporate microwell arrays with sub-10 μm features within a single layer of microfluidic circuitry is presented. Typically, the construction of devices that incorporate very small architecture within larger components has required the assembly of multiple elements to form a working device. Rapid, facile production of a working device using only a single layer of molded polydimethylsiloxane (PDMS) and a glass support substrate is achieved with the reported fabrication technique. A combination of conventional wet-chemical etching for larger (≥20 μm) microchannel features and focused ion beam (FIB) milling for smaller (≤10 μm) microwell features was used to fabricate a monolithic glass master mold. PDMS/glass hybrid chips were then produced using simple molding and oxygen plasma bonding methods. Microwell structures were loaded with 3 μm antibody-functionalized dye-encoded polystyrene spheres, and a sandwich immunoassay for common cytokines was performed to demonstrate proof-of-principle. Potential applications for this device include highly parallel multiplexed sandwich immunoassays, DNA/RNA hybridization analyses, and enzyme linked immunosorbent assay (ELISA). The fabrication technique described can be used for rapid prototyping of devices wherever submicrometer- to micrometer-sized features are incorporated into a microfluidic device.     

Highly elastic conductive polymeric MEMS

Polymeric structures with integrated, functional microelectrical mechanical systems (MEMS) elements are increasingly important in various applications such as biomedical systems or wearable smart devices. These applications require highly flexible and elastic polymers with good conductivity, which can be embedded into a matrix that undergoes large deformations. Conductive polydimethylsiloxane (PDMS) is a suitable candidate but is still challenging to fabricate. Conductivity is achieved by filling a nonconductive PDMS matrix with conductive particles. In this work, we present an approach that uses new mixing techniques to fabricate conductive PDMS with different fillers such as carbon black, silver particles, and multiwalled carbon nanotubes. Additionally, the electrical properties of all three composites are examined under continuous mechanical stress. Furthermore, we present a novel, low-cost, simple three-step molding process that transfers a micro patterned silicon master into a polystyrene (PS) polytetrafluoroethylene (PTFE) replica with improved release features. This PS/PTFE mold is used for subsequent structuring of conductive PDMS with high accuracy. The non sticking characteristics enable the fabrication of delicate structures using a very soft PDMS, which is usually hard to release from conventional molds. Moreover, the process can also be applied to polyurethanes and various other material combinations.     

Fabrication of topologically complex three-dimensional microfluidic systems in PDMS by rapid prototyping

This paper describes a procedure for making topologically complex three-dimensional microfluidic channel systems in poly(dimethylsiloxane) (PDMS). This procedure is called the "membrane sandwich" method to suggest the structure of the final system: a thin membrane having channel structures molded on each face (and with connections between the faces) sandwiched between two thicker, flat slabs that provide structural support. Two "masters" are fabricated by rapid prototyping using two-level photolithography and replica molding. They are aligned face to face, under pressure, with PDMS prepolymer between them. The PDMS is cured thermally. The masters have complementary alignment tracks, so registration is straightforward. The resulting, thin PDMS membrane can be transferred and sealed to another membrane or slab of PDMS by a sequence of steps in which the two masters are removed one at a time; these steps take place without distortion of the features. This method can fabricate a membrane containing a channel that crosses over and under itself, but does not intersect itself and, therefore, can be fabricated in the form of any knot. It follows that this method can generate topologically complex microfluidic systems; this capability is demonstrated by the fabrication of a "basketweave" structure. By filling the channels and removing the membrane, complex microstructures can be made. Stacking and sealing more than one membrane allows even more complicated geometries than are possible in one membrane. A square coiled channel that surrounds, but does not connect to, a straight channel illustrates this type of complexity.     

Highly elastic conductive polymeric MEMS

Abstract

Polymeric structures with integrated, functional microelectrical mechanical systems (MEMS) elements are increasingly important in various applications such as biomedical systems or wearable smart devices. These applications require highly flexible and elastic polymers with good conductivity, which can be embedded into a matrix that undergoes large deformations. Conductive polydimethylsiloxane (PDMS) is a suitable candidate but is still challenging to fabricate. Conductivity is achieved by filling a nonconductive PDMS matrix with conductive particles. In this work, we present an approach that uses new mixing techniques to fabricate conductive PDMS with different fillers such as carbon black, silver particles, and multiwalled carbon nanotubes. Additionally, the electrical properties of all three composites are examined under continuous mechanical stress. Furthermore, we present a novel, low-cost, simple three-step molding process that transfers a micro patterned silicon master into a polystyrene (PS) polytetrafluoroethylene (PTFE) replica with improved release features. This PS/PTFE mold is used for subsequent structuring of conductive PDMS with high accuracy. The non sticking characteristics enable the fabrication of delicate structures using a very soft PDMS, which is usually hard to release from conventional molds. Moreover, the process can also be applied to polyurethanes and various other material combinations.     

ELECTROFORMING TECHNIQUES IN THE LIGA PROCESS FOR THE PRODUCTION OF MICRODEVICES

At the Karlsruhe Nuclear Research Center a new technology, called LIGA process, is being developed. In this process, synchrotron radiation lithography, electroforming and plastic molding are employed for the fabrication of microdevices with characteristic dimensions of few micrometers and structural heights of several hundred micrometers. Besides the production of metallic microdevices made of nickel, copper or gold, electroforming techniques are also used in fabricating X-ray masks and mold inserts to be utilized in the process. Plating processes used In the LIGA process are being automated for integration in a new clean-room plating machine developed for the mass production of microdevices.     

微流控芯片基片与盖片一体化注塑成型研究

针对微流控芯片基片与盖片的结构特点,提出了定模先行抽芯机构,设计制造了微流控芯片基片与盖片一体化注塑成型模具,并进行注塑成型试验研究.结果表明:定模先行抽芯机构可以有效解决盖片上圆孔状储液池成型与脱模的技术难题,如何使微通道复制完全是微流控芯片基片注塑成型的主要技术难点;模具温度对提高微通道复制度起决定性作用,注射速度和熔体温度是次要因素,而注射压力相对其他因素影响力较差,但必须保持在一个较高的水平,依此形成塑料微流控芯片的注塑成型工艺规范.     

Integrated capillary electrophoresis on flexible silicone microdevices: analysis of DNA restriction fragments and detection of single DNA molecules on microchips

Microchips for integrated capillary electrophoresis systems were produced by molding a poly(dimethylsiloxane) (PDMS) silicone elastomer against a microfabricated master. The good adhesion of the PDMS devices on clean planar surfaces allows for a simple and inexpensive generation of networks of sealed microchannels, thus removing the constraints of elaborate bonding procedures. The performance of the devices is demonstrated with both fast separations of φX-174/HaeIII DNA restriction fragments labeled with the intercalating dye YOYO-1 and fluorescently labeled peptides. Detection limits in the order of a few zeptomoles (10(-)(21) mol) have been achieved for each injected DNA fragment, corresponding to a mass detection limit of ～2 fg for the 603 base pair fragment. Single λ-DNA molecules intercalated with YOYO-1 at a base pair-to-dye ratio of 10:1 could be detected with an uncomplicated laser-induced fluorescence detection setup. High single-molecule detection efficiency (>50%) was achieved under electrophoretically controlled mass transport conditions in PDMS microchannels.     

A soft lithographic approach to fabricate patterned microfluidic channels

The control of surface properties and spatial presentation of functional molecules within a microfluidic channel is important for the development of diagnostic assays and microreactors and for performing fundamental studies of cell biology and fluid mechanics. Here, we present a simple technique, applicable to many soft lithographic methods, to fabricate robust microchannels with precise control over the spatial properties of the substrate. In this approach, the patterned regions were protected from oxygen plasma by controlling the dimensions of the poly(dimethylsiloxane) (PDMS) stamp and by leaving the stamp in place during the plasma treatment process. The PDMS stamp was then removed, and the microfluidic mold was irreversibly bonded to the substrate. The approach was used to pattern a nonbiofouling poly(ethylene glycol)-based copolymer or the polysaccharide hyaluronic acid within microfluidic channels. These nonbiofouling patterns were then used to fabricate arrays of fibronectin and bovine serum albumin as well as mammalian cells. In addition, further control over the deposition of multiple proteins onto multiple or individual patterns was achieved using laminar flow. Also, cells that were patterned within channels remained viable and capable of performing intracellular reactions and could be potentially lysed for analysis.     

Cover Picture: Fabrication of Elastomeric Wires by Selective Electroless Metallization of Poly(dimethylsiloxane) (Adv. Mater. 1/2008)

Tricia Carmichael and co‐workers employ a simple, low‐cost method for the fabrication of patterned metal films on elastomeric poly(dimethylsiloxane) (PDMS) substrates, as described on p. 59. The metal/PDMS composites are electrically conductive and mechanically flexible, making them suitable for use in the fabrication of lightweight, flexible devices such as wearable electronics, biocompatible sensors, and artificial nerves, skins, and muscles. Copper wires on PDMS remain conductive when subjected to linear strains of up to 52 %. The utility of these wires is demonstrated by using them as laminated top contacts in an organic light‐emitting device.     

基于PDMS纳米粒子改性的热模压技术

介绍了一种采用纳米SiO2、TiO2改性聚二甲基硅氧烷(PDMS)的方法,研究了两种纳米材料SiO2、TiO2添加的比例对热压效果的影响及优化条件,并分析了其改性后的热膨胀系数及杨氏模量的变化,以及其改性后热压效果的提高进行了初步分析,最后用改性的PDMS材料,快速成型来制造热压模具,热压制作聚甲基丙烯酸甲酯(PMMA)微流控芯片。与常用的金属模具(如镍模具)相比,此方法具有脱模容易,工艺周期短,难度低,重复性好,价格低等优点。     

Fabricating large arrays of microwells with arbitrary dimensions and filling them using discontinuous dewetting

This paper describes the fabrication of large (up to 45 cm(2)) arrays of microwells, with volumes as small as ～3 fL/well and densities as high as 10(7) wells/cm(2). These arrays of microwells are formed by casting an elastomer, poly(dimethylsiloxane) (PDMS), against "masters" prepared by photolithography; arrays of microwells in other polymers can be formed by using a master consisting of posts in PDMS. A straightforward technique, discontinuous dewetting, allows wells to be filled rapidly (typically on the order of 10(4) wells/s) and uniformly with a wide range of liquids. Several rudimentary strategies for addressing microwells are investigated, including electroosmotic pumping and gaseous diffusion.     

Desktop‐Stereolithography 3D‐Printing of a Poly(dimethylsiloxane)‐Based Material with Sylgard‐184 Properties

The advantageous physiochemical properties of poly(dimethylsiloxane) (PDMS) have made it an extremely useful material for prototyping in various technological, scientific, and clinical areas. However, PDMS molding is a manual procedure and requires tedious assembly steps, especially for 3D designs, thereby limiting its access and usability. On the other hand, automated digital manufacturing processes such as stereolithography (SL) enable true 3D design and fabrication. Here the formulation, characterization, and SL application of a 3D‐printable PDMS resin (3DP‐PDMS) based on commercially available PDMS‐methacrylate macromers, a high‐efficiency photoinitiator and a high‐absorbance photosensitizer, is reported. Using a desktop SL‐printer, optically transparent submillimeter structures and microfluidic channels are demonstrated. An optimized blend of PDMS‐methacrylate macromers is also used to SL‐print structures with mechanical properties similar to conventional thermally cured PDMS (Sylgard‐184). Furthermore, it is shown that SL‐printed 3DP‐PDMS substrates can be rendered suitable for mammalian cell culture. The 3DP‐PDMS resin enables assembly‐free, automated, digital manufacturing of PDMS, which should facilitate the prototyping of devices for microfluidics, organ‐on‐chip platforms, soft robotics, flexible electronics, and sensors, among others.     

Fabrication of integrated microelectrodes for electrochemical detection on electrophoresis microchip by electroless deposition and micromolding in capillary technique

A new method for the fabrication of an integrated microelectrode for electrochemical detection (ECD) on an electrophoresis microchip is described. The pattern of the microelectrode was directly made on the surface of a microscope slide through an electroless deposition procedure. The surface of the slide was first selectively coated with a thin layer of sodium silicate through a micromolding in capillary technique provided by a poly(dimethylsiloxane) (PDMS) microchannel; this left a rough patterned area for the anchoring of catalytic particles. A metal layer was deposited on the pattern guided by these catalytic particles and was used as the working electrode. Factors influencing the fabrication procedure were discussed. The whole chip was built by reversibly sealing the slide to another PDMS layer with electrophoresis microchannels at room temperature. This approach eliminates the need of clean room facilities and expensive apparatus such as for vacuum deposition or sputtering and makes it possible to produce patterned electrodes suitable for ECD on microchip under ordinary chemistry laboratory conditions. Also once the micropattern is ready, it allows the researchers to rebuild the electrode in a short period of time when an electrode failure occurs. Copper and gold microelectrodes were fabricated by this technique. Glucose, dopamine, and catechol as model analytes were tested.     

Stereolithography in tissue engineering

Several recent research efforts have focused on use of computer-aided additive fabrication technologies, commonly referred to as additive manufacturing, rapid prototyping, solid freeform fabrication, or three-dimensional printing technologies, to create structures for tissue engineering. For example, scaffolds for tissue engineering may be processed using rapid prototyping technologies, which serve as matrices for cell ingrowth, vascularization, as well as transport of nutrients and waste. Stereolithography is a photopolymerization-based rapid prototyping technology that involves computer-driven and spatially controlled irradiation of liquid resin. This technology enables structures with precise microscale features to be prepared directly from a computer model. In this review, use of stereolithography for processing trimethylene carbonate, polycaprolactone, and poly(d,l-lactide) poly(propylene fumarate)-based materials is considered. In addition, incorporation of bioceramic fillers for fabrication of bioceramic scaffolds is reviewed. Use of stereolithography for processing of patient-specific implantable scaffolds is also discussed. In addition, use of photopolymerization-based rapid prototyping technology, known as two-photon polymerization, for production of tissue engineering scaffolds with smaller features than conventional stereolithography technology is considered.     

Micro- and nanostripes of self-assembled Au nanocrystal superlattices by direct micromolding

A simple, inexpensive direct micromolding method for patterning Au nanocrystal superlattices using a polydimethylsiloxane (PDMS) stamp has been developed. The method involves in situ synthesis of Au(I) dodecanethiolate and its decomposition leading to Au nanocrystals in the microchannels of the stamp which order themselves to form patterned superlattice stripes, in conformity with the stamp geometry. Owing to its insolubility in common solvents, the dodecanethiolate was made by reacting Au(PPh₃)Cl and dodecanethiol in situ inside the microchannels, by injecting first the former solution in toluene at room temperature followed by the thiol solution at 120 °C. Annealing the reaction mixture at 250 °C, resulted in formation of nanocrystals (with a mean diameter of 7.5 nm) and hexagonal ordering. By using an external pressure while molding, parallel stripes with sub-100 nm widths were obtained. The choice of parameters such as injection temperature of the thiol and concentrations is shown to be important if an ordered superlattice is to be obtained. In addition, these parameters can be varied as a means to control the nanocrystal size.     

Fast patterning of poly(methyl methacrylate) by a novel soft molding approach and its application to the fabrication of silver structures

A novel single-step approach for the fabrication of poly(methyl methacrylate) structures by soft molding of a 5 wt% solution in acetone is reported. The use of a low weight solution and of a solvent with high volatility ensures a very fast patterning, down to 10 s. In addition, the process is extremely simple and cost-effective, since just one elastomeric mold is needed, and areas as large as 1 cm² were patterned uniformly and defect-free. The process was applied to the fabrication of silver structures by silver deposition via electroless plating or evaporation followed by poly(methyl methacrylate) removal. Structures of various shapes and sizes, with dimensions in the micrometer and submicrometer range were successfully fabricated, showing the versatility of the process. This silver patterning process is particularly well suited for applications in microelectronics and optoelectronics, such as the fabrication of transparent electrodes for solar cells and displays, manufacturing of metal etching masks and wiring of printed circuits.     

Creating addressable aqueous microcompartments above solid supported phospholipid bilayers using lithographically patterned poly(dimethylsiloxane) molds

Herein we report the use of microcontact displacement (microCD) to generate addressable arrays of aqueous solutions above fluid lipid bilayers by bringing a patterned PDMS mold into conformal contact with a phospholipid membrane on a solid supported substrate. Epifluorescence microscopy established that the bilayer material was displaced in regions where contact between the mold and substrate was made. Photobleaching experiments confirmed that the bilayer sectors remained individually fluid but completely separated. The microcompartments created by microCD could be individually injected with aqueous solutions that remained sealed from their neighbors. This procedure was then exploited for screening a small library of aqueous-phase molecules for their ability to inhibit binding between surface-bound ligands and soluble protein receptors.     

Surface modification of poly(dimethylsiloxane) microfluidic devices by ultraviolet polymer grafting

Poly(dimethylsiloxane) (PDMS)-based microfluidic devices are increasing in popularity due to their ease of fabrication and low costs. Despite this, there is a tremendous need for strategies to rapidly and easily tailor the surface properties of these devices. We demonstrate a one-step procedure to covalently link polymers to the surface of PDMS microchannels by ultraviolet graft polymerization. Acrylic acid, acrylamide, dimethylacrylamide, 2-hydroxylethyl acrylate, and poly(ethylene glycol)monomethoxyl acrylate were grafted onto PDMS to yield hydrophilic surfaces. Water droplets possessed contact angles as low as 45 degrees on the grafted surfaces. Microchannels constructed from the grafted PDMS were readily filled with aqueous solutions in contrast to devices composed of native PDMS. The grafted surfaces also displayed a substantially reduced adsorption of two test peptides compared to that of oxidized PDMS. Microchannels with grafted surfaces exhibited electroosmotic mobilities intermediate to those displayed by native and oxidized PDMS. Unlike the electroosmotic mobility of oxidized PDMS, the electroosmotic mobility of the grafted surfaces remained stable upon exposure to air. The electrophoretic resolution of two test peptides in the grafted microchannels was considerably improved compared to that in microchannels composed of oxidized PDMS. By using the appropriate monomer, it should be possible to use UV grafting to impart a variety of surface properties to PDMS microfluidics devices.