Influence of master fabrication techniques on the characteristics of embossed microfluidic channels

We describe protocols for the fabrication of microfluidic devices in plastics using a number of different embossing masters. Masters were fabricated by deep reactive ion etching (DRIE) of silicon (100), wet etching of silicon (100) and (110), and SU-8 processing. Structures embossed into a cyclo-olefin polymer were characterized in terms of the quality of pattern transfer as well as of the surface roughness. High quality pattern transfer was achieved with masters containing structures with angled sidewalls. Pattern distortions occurring during de-embossing were minimized by using masters consisting of SU-8 (which has a thermal expansion coefficient close to that of the substrates). Structures embossed with SU-8 masters also exhibited the lowest surface roughness. However, due to structural deformation, the reusability of the masters prepared for this study extended to only five embossing experiments. Masters fabricated on silicon, on the other hand, were more robust, but were subject to breakage during the de-embossing phase of the experiment. The results of this study will guide researchers in choosing master fabrication methods that will provide profile and surface characteristics of embossed microfluidic channels that are advantageous to their specific application.     

Surface modification of SU-8 for enhanced biofunctionality and nonfouling properties

SU-8 is a chemically amplified, epoxy-based negative photoresist typically used for producing ultrathick resist layers during device manufacturing in the semiconductor industry. As a simple resist, SU-8 has garnered attention as a possible material for a variety of biomedical applications, including tissue engineering, drug delivery, as well as cell-based screening and sensing. However, as a hydrophobic material, the use of SU-8 is limited due to a high degree of nonspecific adsorption of biomolecules, as well as limited cell attachment. In this work, surface chemistry is utilized to modify the SU-8 surface by covalently attaching poly(ethylene glycol) (PEG) to increase biofunctionality and improve its nonfouling properties. Different molecular weights and concentrations of PEG were used to form films of various grafting densities on SU-8 surfaces. X-ray photoelectron spectroscopy (XPS) was used to verify the presence of PEG moieties on the SU-8 surface. High-resolution C1s spectra show that, with an increase in concentration and immobilization time, the grafting density of PEG also increases. Further, a standard overlayer model was used to calculate the thickness of the PEG films formed. The effect of PEG-modified SU-8 was examined in terms of protein adsorption on the surface and fibroblast-surface interactions.     

Effect of exposure dose on the replication fidelity and profile of very high aspect ratio microchannels in SU-8

We are interested in using SU-8 dense gratings with very high aspect ratio microchannels as the master mold for fabrication of child molds needed for replication. For such applications, the sidewall taper angle and mask replication fidelity of SU-8 are very important. Increasing the exposure time was experimentally observed to decrease the width of the microchannel and the sidewall angle of SU-8 bars. A new diffraction-refraction-reflection model was also developed. The calculated microchannel width and sidewall angle at high exposure dose agreed well with the experimentally observed values indicating that reflection at the silicon substrate was significant. The larger than calculated actual microchannel width for low exposure dose was shown to be due to leaching of unreacted SU-8 in the developer. Dense gratings of high aspect ratio SU-8 bars separated by high aspect ratio (19.1) microchannels were also demonstrated.     

High intensity light emitting diode array as an alternative exposure source for the fabrication of electrophoretic microfluidic devices

A commercially available array of light emitting diodes (LEDs), namely a UV Shark series LED high flux array, was evaluated as a light source for photolithographic patterning of SU-8 photoresist for the fabrication of templates suitable to make poly(dimethylsiloxane) (PDMS) microchips for electrophoresis. At a distance of 15 cm from the substrate, a relatively even intensity of 0.76+/-0.05 mW/cm(2) was obtained over an area sufficient for patterning a 10 cm (4 in.) silicon wafer. The exposure source was evaluated using a spiral mask design covering a 10 cm wafer. PDMS replicates of this template made in a 25 microm thick layer of SU-8 3025 showed little variation in width over the surface of the substrate, with a variation of 3.2% RSD (n=36) and a maximum range in widths of 7.8% of the mean channel width. The use of the optional metal reflector available with the LED array provided partial collimation of the light allowing near vertical structures to be produced across the entire wafer, something which was not possible without the reflector. SU-8 masters prepared using the LED array were compared to masters made using an alternative cheap lithographic source, namely a gel crosslinker. The SU-8 features were much narrower with the LED array than the crosslinker due to the multiple light sources in the crosslinker. A PDMS microchip made using a SU-8 template created using the Shark UV LED array was used for the electrophoretic separation of three anionic fluorescent dyes, with efficiencies up to 32,000 plates. Given that the LED array can be purchased and assembled for less than US$ 500, the Shark UV LED array is a promising alternative to more expensive lithographic light sources and will have significant appeal to many researchers wishing to undertake research in microfluidics around the world.     

Magnetohydrodynamic effects on a charged colloidal sphere with arbitrary double-layer thickness

An analytical study is presented for the magnetohydrodynamic (MHD) effects on a translating and rotating colloidal sphere in an arbitrary electrolyte solution prescribed with a general flow field and a uniform magnetic field at a steady state. The electric double layer surrounding the charged particle may have an arbitrary thickness relative to the particle radius. Through the use of a simple perturbation method, the Stokes equations modified with an electric force term, including the Lorentz force contribution, are dealt by using a generalized reciprocal theorem. Using the equilibrium double-layer potential distribution from solving the linearized Poisson-Boltzmann equation, we obtain closed-form formulas for the translational and angular velocities of the spherical particle induced by the MHD effects to the leading order. It is found that the MHD effects on the particle movement associated with the translation and rotation of the particle and the ambient fluid are monotonically increasing functions of κa, where κ is the Debye screening parameter and a is the particle radius. Any pure rotational Stokes flow of the electrolyte solution in the presence of the magnetic field exerts no MHD effect on the particle directly in the case of a very thick double layer (κa→0). The MHD effect caused by the pure straining flow of the electrolyte solution can drive the particle to rotate, but it makes no contribution to the translation of the particle.     

Development of a multireactor microfluidic system for the determination of DNA using real-time polymerase chain reaction

A microfluidic system that allowed us to perform the real-time polymerase chain reaction (PCR) in a glass-silicon microchip containing nine 250-nL microreactors was developed and studied. The resulting high heating/cooling rates of a PCR mixture in a microreactor allowed us to optimize the amplification mode (1 min/cycle). The silicon surface of microreactors was successfully passivated. The resulting analytical system allowed us to measure the PCR kinetic curves in chip microreactors at a DNA concentration of ～5 × 10⁴ copies per microreactor. It was found that, if the PCR is performed in a microchip with real-time detection using the optimized amplification mode, the result can be obtained 13–14 min after the onset of reaction.     

Reactions of Difunctional Electrophiles with Functionalized Aryllithium Compounds: Remarkable Chemoselectivity by Flash Chemistry

Flash chemistry using flow microreactors enables highly chemoselective reactions of difunctional electrophiles with functionalized aryllithium compounds by virtue of extremely fast micromixing. The approach serves as a powerful method for protecting‐group‐free synthesis using organolithium compounds and opens a new possibility in the synthesis of polyfunctional organic molecules.     

Fabrication and Packaging Technologies of Love-wave-based Microbalance for Fluid Analysis

Quartz crystal microbalance (QCM) based techniques have been developed for years to address various kinds of biochemical analyses in liquid media. An alternative to this approach based on guided acoustic shear waves, the socalled Love wave devices, has been proved to allow for increasing gravimetric sensitivity. However, this approach reveals more complicated to implement as the surface on which reactions are achieved is the same as the one used for electrical connection. As a consequence, a microfluidic set-up must be implemented to prevent unwanted interactions between the corresponding areas (IDTs and propagation path). The main issue when using SAW Sensors for in-liquid biochemical analyses [1-4], especially in a commercial objective, is the development of a reliable and reproducible fluidic system [5] meeting the main following requirements: i) low acoustic leakage. ii) chemically inert to biological samples. iii) reproducible fabrication at the wafer scale level.In the present work we explore the use of the SU-8 epoxy-based photoresist combined with silicon or quartz machined covers for the fabrication of this fluidic circuit. A first structure is fabricated using deep etch lithography, the cover is then glued to the remaining SU-8 structure using a thin glue layer. The packaging system prevents covering the IDTs with liquids and defines the sensing area in the region in-between the IDTs. Once the fabrication achieved, we evaluate the velocity and propagation loss using a network analyzer to measure the influence of the proposed packaging approaches on the principal wave characteristics.     

Surface assisted laser desorption/ionization on two-layered amorphous silicon coated hybrid nanostructures

Matrix-free laser desorption/ionization was studied on two-layered sample plates consisting of a substrate and a thin film coating. The effect of the substrate material was studied by depositing thin films of amorphous silicon on top of silicon, silica, polymeric photoresist SU-8, and an inorganic-organic hybrid. Des-arg⁹-bradykinin signal intensity was used to evaluate the sample plates. Silica and hybrid substrates were found to give superior signals compared with silicon and SU-8 because of thermal insulation and compatibility with amorphous silicon deposition process. The effect of surface topography was studied by growing amorphous silicon on hybrid micro- and nanostructures, as well as planar hybrid. Compared with planar sample plates, micro- and nanostructures gave weaker and stronger signals, respectively. Different coating materials were tested by growing different thin film coatings on the same substrate. Good signals were obtained from titania and amorphous silicon coated sample plates, but not from alumina coated, silicon nitride coated, or uncoated sample plates. Overall, the strongest signals were obtained from oxygen plasma treated and amorphous silicon coated inorganic-organic hybrid, which was tested for peptide-, protein-, and drug molecule analysis. Peptides and drugs were analyzed with little interference at low masses, subfemtomole detection levels were achieved for des-arg⁹-bradykinin, and the sample plates were also suitable for ionization of small proteins.     

An open-framework silicogermanate with 26-ring channels built from seven-coordinated (Ge,Si)10(O,OH)28 clusters

We report a new open-framework silicogermanate SU-61 containing 26-ring channels with a low framework density. It can be seen as a crystalline analogue to the mesoporous silica MCM-41. The structure is built from the assembly of (Ge,Si)10(O,OH)28 clusters. It is the first time that silicon has been successfully introduced in the Ge10 cluster in significant amounts ( approximately 21% of the tetrahedral sites). Five- and six-coordinated Ge10 clusters have previously been observed in other germanate compounds leading to either dense or open structures. In SU-61, the seven-coordinated clusters fall onto yet another underlying net, the osf net. SU-61, along with other Ge10 based frameworks, shows the versatility of the germanate system to adopt defined topologies playing on the connectivity of the clusters following the principles of net decoration and scale chemistry.     

Immobilized enzyme studies in a microscale bioreactor

Novel microreactors with immobilized enzymes were fabricated using both silicon and polymer-based microfabrication techniques. The effectiveness of these reactors was examined along with their behavior over time. Urease enzyme was successfully incorporated into microchannels of a polymeric matrix of polydimethylsiloxane and through layer-bylayer self-assembly techniques onto silicon. The fabricated microchannels had cross-sectional dimensions ranging from tens to hundreds of micrometers in width and height. The experimental results for continuous-flow microreactors are reported for the conversion of urea to ammonia by urease enzyme. Urea conversions of > 90% were observed.     

Simple photografting method to chemically modify and micropattern the surface of SU-8 photoresist

SU-8 has gained widespread acceptance as a negative photoresist. It is also finding increasing use as a structural material in microanalytical devices. Consequently, methods to tailor the surface properties of SU-8 as well as to micropattern coatings on the surface of SU-8 are needed. The SU-8 photoresist consists of EPON SU-8 resin mixed with the photoacid generator triarylsulfonium hexafluoroantimonate. This photoacid generator can also serve as a photoinitiator generating free radicals when illuminated with UV light. Under the appropriate conditions, sufficient triarylsulfonium hexafluoroantimonate remains within cured SU-8 to act as a source of free radicals and initiate UV-mediated grafting of polymers onto the surface of the SU-8. UV-mediated grafting was used to coat SU-8 surfaces with poly(acrylic acid) and other water-soluble monomers. The SU-8 surface was chemically micropatterned by placing a mask between the UV light and SU-8. The X-Y spatial resolution of micropatterned poly(acrylic acid) on the SU-8 surface was 2 mum. Three applications of these chemically modified SU-8 surfaces were demonstrated. In the first, poly(ethylene glycol) was used to protect the SU-8 from interactions with proteins, yielding a surface resistant to biofouling. In the second demonstration, the SU-8 surface was micropatterned with a cell-resistant layer to guide cellular attachment and growth. In the final application, SU-8 micropallets were encoded with polymer lines. The bar codes were read by either absorbance or fluorescence measurements. Thus, UV-mediated graft polymerization is an efficient and effective method to micropattern coatings onto the surface of SU-8.     

Oligosaccharide synthesis in microreactors

Described is the combination of microreactors and fluorous phase chemistry to assemble oligosaccharides. The synthesis of a beta-(1-->6) linked D-glucopyranoside homotetramer serves to illustrate this approach. Glycosylations employing a Fmoc-protected glucosyl phosphate building block were performed in a silicon-based micro-structured device to optimize reaction conditions and for reaction scale-up. A perfluorinated linker at the reducing end of the oligosaccharides allowed for purification by fluorous solid-phase extraction (FSPE) and further functionalization.     

Flash chemistry: fast chemical synthesis by using microreactors

This concept article provides a brief outline of the concept of flash chemistry for carrying out extremely fast reactions in organic synthesis by using microreactors. Generation of highly reactive species is one of the key elements of flash chemistry. Another important element of flash chemistry is the control of extremely fast reactions to obtain the desired products selectively. Fast reactions are usually highly exothermic, and heat removal is an important factor in controlling such reactions. Heat transfer occurs very rapidly in microreactors by virtue of a large surface area per unit volume, making precise temperature control possible. Fast reactions often involve highly unstable intermediates, which decompose very quickly, making reaction control difficult. The residence time can be greatly reduced in microreactors, and this feature is quite effective in controlling such reactions. For extremely fast reactions, kinetics often cannot be used because of the lack of homogeneity of the reaction environment when they are conducted in conventional reactors such as flasks. Fast mixing using micromixers solves such problems. The concept of flash chemistry has been successfully applied to various organic reactions including a) highly exothermic reactions that are difficult to control in conventional reactors, b) reactions in which a reactive intermediate easily decomposes in conventional reactors, c) reactions in which undesired byproducts are produced in the subsequent reactions in conventional reactors, and d) reactions whose products easily decompose in conventional reactors. The concept of flash chemistry can be also applied to polymer synthesis. Cationic polymerization can be conducted with an excellent level of molecular-weight control and molecular-weight distribution control.     

Beyond PDMS: off-stoichiometry thiol-ene (OSTE) based soft lithography for rapid prototyping of microfluidic devices

In this article we introduce a novel polymer platform based on off-stoichiometry thiol-enes (OSTEs), aiming to bridge the gap between research prototyping and commercial production of microfluidic devices. The polymers are based on the versatile UV-curable thiol-ene chemistry but takes advantage of off-stoichiometry ratios to enable important features for a prototyping system, such as one-step surface modifications, tuneable mechanical properties and leakage free sealing through direct UV-bonding. The platform exhibits many similarities with PDMS, such as rapid prototyping and uncomplicated processing but can at the same time mirror the mechanical and chemical properties of both PDMS as well as commercial grade thermoplastics. The OSTE-prepolymer can be cast using standard SU-8 on silicon masters and a table-top UV-lamp, the surface modifications are precisely grafted using a stencil mask and the bonding requires only a single UV-exposure. To illustrate the potential of the material we demonstrate key concepts important in microfluidic chip fabrication such as patterned surface modifications for hydrophobic stops, pneumatic valves using UV-lamination of stiff and rubbery materials as well as micromachining of chip-to-world connectors in the OSTE-materials.     

An integrated optical oxygen sensor fabricated using rapid-prototyping techniques

This paper details the design and fabrication of an integrated optical biochemical sensor using a select oxygen-sensitive fluorescent dye, tris(2,2'-bipyridyl) dichlororuthenium(ii) hexahydrate, combined with polymeric waveguides that are fabricated on a glass substrate. The sensor uses evanescent interaction of light confined within the waveguide with the dye that is immobilized on an SU-8 waveguide surface. Adhesion of the dye to the integrated waveguide surface is accomplished using a unique process of spin-coating/electrostatic layer-by-layer formation. The SU-8 waveguide was chemically modified to allow the deposition process. Exposure of the dye molecules to the analyte and subsequent chemical interaction is achieved by directly coupling the fluid channel to the integrated waveguide. The completed sensor was linear in the dissolved oxygen across a wide range of interest and had a sensitivity of 0.6 ppm. A unique fabrication aspect of this sensor is the inherent simplicity of the design, and the resulting rapidity of fabrication, while maintaining a high degree of functionality and flexibility.     

Active 3-D microscaffold system with fluid perfusion for culturing in vitro neuronal networks

This work demonstrated the design, fabrication, packaging, and characterization of an active microscaffold system with fluid perfusion/nutrient delivery functionalities for culturing in vitro neuronal networks from dissociated hippocampal rat pup neurons. The active microscaffold consisted of an 8 x 8 array of hollow, microfabricated, SU-8 towers (1.0 mm or 1.5 mm in height), with integrated, horizontal, SU-8 cross-members that connect adjacent towers, thus forming a 3-D grid that is conducive to branching, growth, and increased network formation of dissociated hippocampal neurons. Each microtower in the microscaffold system contained a hollow channel and multiple fluid ports for media delivery and perfusion of nutrients to the in vitro neuronal network growing within the microscaffold system. Additionally, there were two exposed Au electrodes on the outer wall of each microtower at varying heights (with insulated leads running within the microtower walls), which will later allow for integration of electrical stimulation/recording functionalities into the active microscaffold system. However, characterization of the stimulation/recording electrodes was not included in the scope of this paper. Design, fabrication, fluid packaging, and characterization of the active microscaffold system were performed. Furthermore, use of the active microscaffold system was demonstrated by culturing primary hippocampal embryonic rat pup neurons, and characterizing cell viability within the microscaffold system.     

Development of novel microscale system as immobilized enzyme bioreactor

This study involves a novel method for immobilized enzyme catalysis. The focus of the work was to design and construct a microscale bioreactor using microfabrication techniques traditionally employed within the semiconductor industry. Enzymes have been immobilized on the microreactor walls by incorporating them directly into the wall material. Fabricated microchannels have cross-sectional dimensions on the order of hundreds of micrometers, constructed using polydimethylsiloxane cast on silicon/SU-8 molds. The resulting ratio of high surface area to volume creates an efficient, continuous-flow reaction system. Transverse features also containing enzymes were molded directly into the channels.     

A planar microfabricated nanoelectrospray emitter tip based on a capillary slot

We report on the fabrication and testing of planar nib-like structures for nanoelectrospray ionization-mass spectrometry (nanoESI-MS) applications. The micro-nib structures were fabricated on silicon substrates using the negative photoresist SU-8; they include capillary slots with widths of 8 and 16 microm. A suitable wafer cleaving step made the nib-like structures overhang the edge of a silicon substrate to provide a robust interface for nanoESI-MS applications; this freeing of the nib tip from the wafer surface created a point-like structure that is essential to establish an electrospray. The micro-nib sources were successfully tested on an LCQ Deca XP+ ion trap mass spectrometer using peptide samples at concentrations down to 1 microM. The high voltage was applied using a platinum wire inserted in the sample reservoir upstream to the capillary slot. A Taylor cone was clearly seen at the nib tip. The micro-nibs performed well at voltages as low as 0.8 kV; such performances are state-of-the-art with respect to current micromachined ESI-MS interfaces and are conditions comparable to those used for standard emitter tips. In addition, we clearly observed the influence of the micro-nib slot width on the ionization performances: the narrower the slot, the better the performances.     

Microstencils for the patterning of nontraditional materials

A microfabrication technique that uses a photolithographically patterned film as a microstencil has been developed. This microstencil has a bilayer structure comprised of parylene and SU-8 films with thicknesses from 4 to 100 microm. The parylene layer enables the microstencil to be mechanically peeled from hydrophilic substrates. Since no chemicals are required to release the microstencil, this technique can be used to pattern chemically and biologically sensitive materials. The amount of material deposited can be automatically controlled by the height of the SU-8 structures or externally controlled by spin coating or other thin film deposition techniques. This patterning method is very versatile and has been used to pattern features as small as 25 by 25 microm on silicon, glass, and polymer substrates. As an initial demonstration, we have patterned wax, cells, proteins, sol, and CYTOP.