Biocompatible "click" wafer bonding for microfluidic devices

We introduce a novel dry wafer bonding concept designed for permanent attachment of micromolded polymer structures to surface functionalized silicon substrates. The method, designed for simultaneous fabrication of many lab-on-chip devices, utilizes a chemically reactive polymer microfluidic structure, which rapidly bonds to a functionalized substrate via"click" chemistry reactions. The microfluidic structure consists of an off-stoichiometry thiol-ene (OSTE) polymer with a very high density of surface bound thiol groups and the substrate is a silicon wafer that has been functionalized with common bio-linker molecules. We demonstrate here void free, and low temperature (< 37 °C) bonding of a batch of OSTE microfluidic layers to a silane functionalized silicon wafer.     

Biofunctionalization of electrowetting-on-dielectric digital microfluidic chips for miniaturized cell-based applications

In this paper we report on the controlled biofunctionalization of the hydrophobic layer of electrowetting-on-dielectric (EWOD) based microfluidic chips with the aim to execute (adherent) cell-based assays. The biofunctionalization technique involves a dry lift-off method with an easy to remove Parylene-C mask and allows the creation of spatially controlled micropatches of biomolecules in the Teflon-AF(?) layer of the chip. Compared to conventional methods, this method (i) is fully biocompatible; and (ii) leaves the hydrophobicity of the chip surface unaffected by the fabrication process, which is a crucial feature for digital microfluidic chips. In addition, full control of the geometry and the dimensions of the micropatches is achieved, allowing cells to be arrayed as cell clusters or as single cells on the digital microfluidic chip surface. The dry Parylene-C lift-off technique proves to have great potential for precise biofunctionalization of digital microfluidic chips, and can enhance their use for heterogeneous bio-assays that are of interest in various biomedical applications.     

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.     

Field-effect flow control in a polydimethylsiloxane-based microfluidic system.

The application of the field-effect for direct control of electroosmosis in a polydimethylsiloxane (PDMS)-based microfluidic system, constructed on a silicon wafer with a 2.0 microm electrically insulating layer of silicon dioxide, is demonstrated. This microfluidic system consists of a 2.0 cm open microchannel fabricated on a PDMS slab, which can reversibly adhere to the silicon wafer to form a hybrid microfluidic device. Aside from mechanically serving as a robust bottom substrate to seal the channel and support the microfluidic system, the silicon wafer is exploited to achieve field-effect flow control by grounding the semiconductive silicon medium. When an electric field is applied through the channel, a radial electric potential gradient is created across the silicon dioxide layer that allows for direct control of the zeta potential and the resulting electroosmotic flow (EOF). By configuring this microfluidic system with two power supplies at both ends of the microchannel, the applied electric potentials can be varied for manipulating the polarity and the magnitude of the radial electric potential gradient across the silicon dioxide layer. At the same time, the longitudinal potential gradient through the microchannel, which is used to induce EOF, is held constant. The results of EOF control in this hybrid microfluidic system are presented for phosphate buffer at pH 3 and pH 5. It is also demonstrated that EOF control can be performed at higher solution pH of 6 and 7.4 by modifying the silicon wafer surface with cetyltrimethylammonium bromide (CTAB) prior to assembly of the hybrid microfluidic system. Results of EOF control from this study are compared with those reported in the literature involving the use of other microfluidic devices under comparable solution conditions.     

Fabrication of microfluidic devices using photopatternable hybrid sol-gel coatings

UV-patternable organic-inorganic hybrid sol-gel coating was used to develop microchannel on silicon and glass wafers by photolithography processing. The sol-gel coating was formed with 3-methacryloxypropyltrimethoxysilane (MPTS) as photosensitive component and zirconium propoxide as property modifier. In order to enhance the UV light efficiency during photolithography processing for glass substrates, a thin copper layer with thickness of 100–200 nm was deposited on one side of the glass wafer. The closed microchannels were formed by bonding of two developed surface channels of compensating patterns by wafer bonding technology. The bonding material is the same as the channel body ensuring a uniform surface property for the microchannels. A thin layer of about 2 μm was applied on the developed channels by spin coating. The depth of the surface channels on each wafer is about 10–12 μm, and the height of the closed channels is therefore in the range of 22–24 μm. Different microfluidic devices such as chaotic micromixers and microsplitters were fabricated.     

Multi-layer microfluidic glass chips for microanalytical applications

A new, versatile architecture is presented for microfluidic devices made entirely from glass, for use with reagents which would prove highly corrosive for silicon. Chips consist of three layers of glass wafers bonded together by fusion bonding. On the inside wafer faces a network of microfluidic channels is created by photolithography and wet chemical etching. Low dead-volume fluidic connections between the layers are fabricated by spark-assisted etching (SAE), a computer numerical controlled (CNC)-like machining technique new to microfluidic system fabrication. This method is also used to form a vertical, long path-length, optical cuvette through the middle wafer for optical absorbance detection of low-concentration compounds. Advantages of this technique compared with other, more standard, methods are discussed. When the new glass-based device for flow-injection analysis of ammonia was compared with our first-generation chips based on silicon micromachining, concentration sensitivity was higher, because of the longer path-length of the optical cuvette. The dependence of dispersion on velocity profile and on channel cross-sectional geometry is discussed. The rapid implementation of the devices for an organic synthesis reaction, the Wittig reaction, is also briefly described.     

Fabrication of SU-8 multilayer microstructures based on successive CMOS compatible adhesive bonding and releasing steps

This paper describes a novel fabrication process based on successive wafer-level bonding and releasing steps for stacking several patterned layers of the negative photoresist EPON SU-8. This work uses a polyimide film to enhance previous low temperature bonding technology. The film acts as a temporary substrate where the SU-8 is photopatterned. The poor adhesion between the polyimide film and SU-8 allows the film to be released after the bonding process, even though the film is still strong enough to carry out photolithography. Using this technique, successive adhesive bonding steps can be carried out to obtain complex 3-D multilayer structures. Interconnected channels with smooth vertical sidewalls and freestanding structures are fabricated. Unlike previous works, all the layers are photopatterned before the bonding process yielding sealed cavities and complex three-dimensional structures without using a sacrificial layer. Adding new SU-8 layers reduces the bonding quality because each additional layer decreases the thickness uniformity and increases the polymer crosslinking level. The effect of these parameters is quantified in this paper. This process guarantees compatibility with CMOS electronics and MEMS. Furthermore, the releasing step leaves the input and the output of the microchannels in contact with the outside world, avoiding the usual slow drilling process of a cover. Hence, in addition to the straightforward integration of electrodes on a chip, this fabrication method facilitates the packaging of these microfluidic devices.     

Nanofluidic channels by anodic bonding of amorphous silicon to glass to study ion-accumulation and ion-depletion effect

A unique phenomenon, ion-enrichment and ion-depletion effect, exists in nanofluidic channels and is observed in amorphous silicon (α-Si) nanochannels as shallow as 50 nm. As a voltage is applied across a nanochannel, ions are rapidly enriched at one end and depleted at the other end of the nanochannel. α-Si is deposited on glass by plasma enhanced chemical vapor deposition and is selectively etched to form nanochannels. The depth of nanochannels is defined by the thickness of the α-Si layer. Low temperature anodic bonding of α-Si to glass was used to seal the channel with a second glass wafer. The strength of the anodic bond was optimized by the introduction of a silicon nitride adhesion promoting layer and double-sided bonding resulting from the electric field reversal. Completed channels, 50 nm in depth, 5 micron wide, and 1 mm long were completely and reliably sealed. Structures based on nanochannels 50-300 nm deep were successfully incorporated into nanofluidic devices to investigate ionic accumulation and depletion effect due to overlapping of electric double layer.     

A novel metal-protected plasma treatment for the robust bonding of polydimethylsiloxane

We describe a method for the irreversible bonding of PDMS-based microfluidic components by exploiting the first reported "shelfable" plasma treatment of PDMS. Simultaneous plasma activation and protection of PDMS surfaces are achieved via RF magnetron sputtering of thin aluminium films in the presence of an argon plasma. In this process, Ar plasma exposure generates a hydrophilic, silanol-enriched polymer surface amenable to irreversible bonding to glass, PDMS or silicon substrates, while the aluminium film functions as a capping layer to preserve the surface functionality over several weeks of storage in ambient conditions. Prior to bonding, this protective aluminium layer is removed by immersion in an aqueous etchant, exposing the adhesive surface. Employing this technology, PDMS-glass and PDMS-PDMS microfluidic devices were fabricated and the adhesive strength was quantified by tensile and leakage testing. Bonding success rates in excess of 80% were demonstrated for both PDMS-glass and PDMS-PDMS assemblies sealed 24 h and 7 days following initial polymer surface activation. PDMS-glass microdevices performed optimally, displaying maximum adhesive strengths on the order of 5 MPa and burst flow rates of approximately 1 mL min(-1) (channel dimensions: l = 25 mm; w = 300 microm; h = 20 microm). These data demonstrate a significant improvement in performance over previously reported bonding technologies, resulting in the production of more robust, longer-lasting microfluidic systems that can withstand higher pressures and flow rates.     

Surface Characterisation of Silicon Based MEMS

Polysilicon Microelectromechanical systems (MEMS) are the subject of intensive researches. Surface chemistry and topography of a MEMS test structure fabricated at Sandia National Laboratory, USA, were studied by means of scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and atom-ic force microscopy(AFM). XPS C1, and Si2p spectra from the polysilicon components, silicon nitride sub-strate and a reference silicon wafer were compared. The results confirm the presence of a self-assembled monolayer (SAM) on the MEMS surface. An island-like morphology was found on both polysilicon and sili-con nitride surfaces of the MEMS. The islands take the form of caps, being up to 0. 5 μm in diameter and 20 nm in height. It is concluded that the co-existence of columnar growth and equiaxed growth during the low pressure chemical vapor deposition(LPCVD) of these layers leads to the observed morphology and the is-lands are caps to the columnar structures.     

Facile fabrication of microfluidic systems using electron beam lithography

We present two fast and generic methods for the fabrication of polymeric microfluidic systems using electron beam lithography: one that employs spatially varying electron-beam energy to expose to different depths a negative electron-beam resist, and another that employs a spatially varying electron-beam dose to differentially expose a bi-layer resist structure. Using these methods, we demonstrate the fabrication of various microfluidic unit structures such as microchannels of a range of geometries and also other more complex structures such as a synthetic gel and a chaotic mixer. These are made without using any separate bonding or sacrificial layer patterning and etching steps. The schemes are inherently simple and scalable, afford high resolution without compromising on speed and allow post CMOS fabrication of microfluidics. We expect them to prove very useful for the rapid prototyping of complete integrated micro/nanofluidic systems with sense and control electronics fabricated by upstream processes.     

Capillary liquid chromatography-microchip atmospheric pressure chemical ionization-mass spectrometry

A miniaturized nebulizer chip for capillary liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (capillary LC-microchip APCI-MS) is presented. The APCI chip consists of two wafers, a silicon wafer and a Pyrex glass wafer. The silicon wafer has a DRIE etched through-wafer nebulizer gas inlet, an edge capillary insertion channel, a stopper, a vaporizer channel and a nozzle. The platinum heater electrode and pads for electrical connection were patterned on to the Pyrex glass wafer. The two wafers were joined by anodic bonding, creating a microchip version of an APCI-source. The sample inlet capillary from an LC column is directly connected to the vaporizer channel of the APCI chip. The etched nozzle in the microchip forms a narrow sample plume, which is ionized by an external corona needle, and the formed ions are analyzed by a mass spectrometer. The nebulizer chip enables for the first time the use of low flow rate separation techniques with APCI-MS. The performance of capillary LC-microchip APCI-MS was tested with selected neurosteroids. The capillary LC-microchip APCI-MS provides quantitative repeatability and good linearity. The limits of detection (LOD) with a signal-to-noise ratio (S/N) of 3 in MS/MS mode for the selected neurosteroids were 20-1000 fmol (10-500 nmol l(-1)). LODs (S/N = 3) with commercial macro APCI with the same compounds using the same MS were about 10 times higher. Fast heat transfer allows the use of the optimized temperature for each compound during an LC run. The microchip APCI-source provides a convenient and easy method to combine capillary LC to any API-MS equipped with an APCI source. The advantages and potentials of the microchip APCI also make it a very attractive interface in microfluidic APCI-MS.     

Low-distortion, high-strength bonding of thermoplastic microfluidic devices employing case-II diffusion-mediated permeant activation

We demonstrate a new method for joining thermoplastic surfaces to produce microfluidic devices. The method takes advantage of the sharply defined permeation boundary of case-II diffusion to generate dimensionally controlled, activated bonding layers at the surfaces being joined. The technique is capable of producing bonds that exhibit cohesive failure, while preserving the fidelity of fine features in the bonding interface. This approach is uniquely suited to production of layered microfluidic structures, as it allows the bond-forming interface between plastic parts to be precisely manipulated at micrometre length scales. Distortions in microfluidic device channels are limited to the size scale of the permeant-swollen layer; 6 microm deep channels are routinely produced with no detectable cross-sectional distortions. Conventional thermal diffusion bonding of identical parts yields less strongly bonded microfluidic structures with increasingly severe dimensional compressions as bonding temperatures approach the thermoplastic glass-transition temperature: a preliminary rheological analysis is consistent with the observed compressions. The bond-enhancing procedure is easily integrated in standard process flows, uses inexpensive reagents, and requires no specialized equipment.     

CO2 laser microfabrication of an integrated polymer microfluidic manifold for the determination of phosphorus

A simple colorimetric technique is implemented in a polymer microfluidic manifold. The simple chemistry aids an uncomplicated microchannel design, which is fabricated by CO(2) laser ablation. Issues such as bonding of multiple layers, alignment of micro-fabricated structures and integration of optical components are addressed. A demonstration of a stopped flow regime in the microfluidic manifold is also presented.     

Chemically resistant microfluidic valves from Viton® membranes bonded to COC and PMMA

We present a reliable technique for irreversibly bonding chemically inert Viton? membranes to PMMA and COC substrates to produce microfluidic devices with integrated elastomeric structures. Viton? is widely used in commercially available valves and has several advantages when compared to other elastomeric membranes currently utilised in microfluidic valves (e.g. PDMS), such as high solvent resistance, low porosity and high temperature tolerance. The bond strength was sufficient to withstand a fluid pressure of 400 kPa (PMMA/Viton?) and 310 kPa (COC/Viton?) before leakage or burst failure, which is sufficient for most microfluidic applications. We demonstrate and characterise on-chip pneumatic Viton? microvalves on PMMA and COC substrates. We also provide a detailed method for bonding fluorinated Viton? elastomer, a highly chemically compatible material, to PMMA and COC polymers. This allows the production of microfluidic devices able to handle a wide range of chemically harsh fluids and broadens the scope of the microfluidic platform concept.     

Pinched flow fractionation devices for detection of single nucleotide polymorphisms

We demonstrate a new and flexible microfluidic based method for genotyping single nucleotide polymorphisms (SNPs). The method relies on size separation of selectively hybridized polystyrene microspheres in a microfluidic pinched flow fractionation (PFF) device. The microfluidic PFF devices with 13 mum deep channels were fabricated by thermal nanoimprint lithography (NIL) in a thin film of cyclic-olefin copolymer (mr-I T85) on a silicon wafer substrate, and the channels were sealed by thermal polymer bonding. Streptavidin coated polystyrene microspheres with a mean diameter of 3.09 microm and 5.6 microm were functionalized with biotin-labeled oligonucleotides for the detection of a mutant (Mt) or wild-type (Wt) DNA sequence in the HBB gene, respectively. Hybridization to functionalized beads was performed with fluorescent targets comprising synthetic DNA oligonucleotides or amplified RNA, synthesized using human DNA samples from individuals with point mutations in the HBB gene. Following a stringent wash, the beads were separated in a PFF device and the fluorescent signal from the beads was analyzed. Patients being wildtypes, heterozygotes or mutated respectively for the investigated mutation could reliably be diagnosed in the PFF device. This indicates that the PFF technique can be used for accurate and fast genotyping of SNPs.     

Silicon surface modification with a mixed silanes layer to immobilize proteins for biosensor with imaging ellipsometry

One kind of surface modification method on silicon wafer was presented in this paper. A mixed silanes layer was used to modify silicon surface and rendered the surface medium hydrophobic. The mixed silanes layer contained two kinds of compounds, aminopropyltriethoxysilane (APTES) and methyltriethoxysilane (MTES). A few of APTES molecules in the layer was used to immobilize covalently human immunoglobulin G (IgG) on the silicon surface. The human IgG molecules immobilized covalently on the modified surface could retain their structures well and bind more antibody molecules than that on silicon surface modified with only APTES. This kind of surface modification method effectively improved the sensitivity of the biosensor with imaging ellipsometry.     

Rapid, sensitive, and multiplexed on-chip optical sensors for micro-gas chromatography

We developed and characterized a rapid, sensitive and integrated optical vapor sensor array for micro-gas chromatography (μGC) applications. The sensor is based on the Fabry-Pérot (FP) interferometer formed by a micrometre-thin vapor-sensitive polymer layer coated on a silicon wafer. The thickness and the refractive index of the polymer vary in response to the vapor analyte, resulting in a change in the reflected intensity of the laser impinged on the sensor. In our study, four different polymers were coated on four wells pre-etched on a silicon wafer to form a spatially separated sensor array. A CMOS imager was employed to simultaneously monitor the polymers' response, thus enabling multiplexed detection of a vapor analyte passing through the GC column. A sub-second detection time was demonstrated. In addition, a sub-picogram detection limit was achieved, representing orders of magnitude improvement over the on-chip vapor sensors previously reported.     

Solder-based chip-to-tube and chip-to-chip packaging for microfluidic devices

Constructing a microsystem compatible with a large variety of chemistries requires a system design that will be robust in the presence of different compounds and at a wide range of conditions. Although microreactors themselves can accommodate a great span of conditions, few packaging schemes are compatible with cryogenic temperatures, high pressures, and aggressive organic solvents. Solder-based connections are designed and implemented on silicon-based microreactors and are demonstrated to withstand elevated pressures (up to 200 atm), a wide range of temperatures (-78 to 160 degrees C) and a variety of solvent systems. Through the deposition of metal bonding pads directly onto silicon and glass surfaces, solder-based chip-to-tube connections can be reliably formed using handheld soldering tools. Packaging techniques are also described for fluidic chip-to-chip bonds, facilitating direct connection of microfluidic modules. This method greatly expands the utility of microfluidic reactors while enabling easy and reproducible fluidic packaging.     

Design, fabrication and characterization of monolithic embedded parylene microchannels in silicon substrate

This paper presents a novel channel fabrication technology of bulk-micromachined monolithic embedded polymer channels in silicon substrate. The fabrication process favorably obviates the need for sacrificial materials in surface-micromachined channels and wafer-bonding in conventional bulk-micromachined channels. Single-layer-deposited parylene C (poly-para-xylylene C) is selected as a structural material in the microfabricated channels/columns to conduct life science research. High pressure capacity can be obtained in these channels by the assistance of silicon substrate support to meet the needs of high-pressure loading conditions in microfluidic applications. The fabrication technology is completely compatible with further lithographic CMOS/MEMS processes, which enables the fabricated embedded structures to be totally integrated with on-chip micro/nano-sensors/actuators/structures for miniaturized lab-on-a-chip systems. An exemplary process was described to show the feasibility of combining bulk micromachining and surface micromachining techniques in process integration. Embedded channels in versatile cross-section profile designs have been fabricated and characterized to demonstrate their capabilities for various applications. A quasi-hemi-circular-shaped embedded parylene channel has been fabricated and verified to withstand inner pressure loadings higher than 1000 psi without failure for micro-high performance liquid chromatography (microHPLC) analysis. Fabrication of a high-aspect-ratio (internal channel height/internal channel width, greater than 20) quasi-rectangular-shaped embedded parylene channel has also been presented and characterized. Its implementation in a single-mask spiral parylene column longer than 1.1 m in a 3.3 mm x 3.3 mm square size on a chip has been demonstrated for prospective micro-gas chromatography (microGC) and high-density, high-efficiency separations. This proposed monolithic embedded channel technology can be extensively implemented to fabricate microchannels/columns in high-pressure microfluidics and high-performance/high-throughput chip-based micro total analysis systems (microTAS).