Bulk modification of polymeric microfluidic devices

The surface properties of microfluidic devices play an important role in their flow behavior. We report here on an effective control of the surface chemistry and performance of polymeric microchips through a bulk modification route during the fabrication process. The new protocol is based on modification of the bulk microchip material by tailored copolymerization of monomers during atmospheric-pressure molding. A judicious addition of a modifier to the primary monomer solution thus imparts attractive properties to the plastic microchip substrate, including significant enhancement and/or modulation of the EOF (with flow velocities comparable to those of glass), a strong pH sensitivity and high stability. Carboxy, sulfo, and amino moieties have thus been introduced (through the incorporation of methylacrylic acid, 2-sulfoethyl-methacrylate and 2-aminoethyl-methacrylate monomers, respectively). A strong increase in the electroosmotic pumping compared to the native poly(methylmethacrylate)(PMMA) microchip (ca. electroosmotic mobility increases from 2.12 to 4.30 x 10(-4) cm(2) V(-1) s(-1)) is observed using a 6% methylacrylate (MAA) modified PMMA microchip. A 3% aminoethyl modified PMMA microchip exhibits a reversal of the electroosmotic mobility (for example, -5.6 x 10(-4) cm(2) V(-1) s(-1) at pH 3.0). The effects of the modifier loading and the pH on the EOF have been investigated for the MAA-modified PMMA chips. The bulk-modified devices exhibit reproducible and stable EOF behavior. The one step fabrication/modification protocol should further facilitate the widespread production of high-performance plastic microchip devices.     

Electrophoresis microchips with sharp inlet tips, for contactless conductivity detection, fabricated by in-situ surface polymerization

A novel method based on in-situ surface polymerization of methyl methacrylate (MMA) has been developed for rapid fabrication of poly(methyl methacrylate) (PMMA) electrophoresis microchips with sharp inlet tips. Prepolymerized MMA containing an ultraviolet (UV) initiator was directly sandwiched between a nickel template and a PMMA plate. The image of the relief on the nickel template was precisely replicated in the synthesized PMMA layer on the surface of the commercially available PMMA plate during UV-initiated polymerization at room temperature. The chips were subsequently assembled by thermal bonding of channel plates and cover sheets. The sample was directly introduced into the separation channel through a sharp inlet tip, which was placed in the sample vial, without use of an injection cross. The attractive performance of the novel PMMA microchips has been demonstrated by using contactless conductivity detection for determination of several inorganic ions. Such rapid and simple sample introduction leads to highly reproducible signals with relative standard deviations of less than 5% for peak responses. These new approaches significantly simplify the process of fabricating PMMA devices and show great promise for high-speed microchip analysis.      

Fabrication of a gold microelectrode for amperometric detection on a polycarbonate electrophoresis chip by photodirected electroless plating

A novel method of photoresist-free micropatterning coupled with electroless gold plating is described for the fabrication of an integrated gold electrode for electrochemical detection (ED) on a polycarbonate (PC) electrophoresis microchip. The microelectrode layout was photochemically patterned onto the surface of a PC plate by selective exposure of the surface coated without photoresist to 254 nm UV light through a chromium/quartz photomask. Thus, the PC plate was selectively sensitized by formation of reactive chemical moieties in the exposed areas. After a series of wet chemistry reactions, the UV-exposed area was activated with a layer of gold nanoparticles that served as a seed to catalyze the electroless plating. The gold microelectrode was then selectively plated onto the activated area by using an electroless gold plating bath. Nonselective gold deposition on the unwanted areas was eliminated by sonication of the activated PC plate in a KSCN solution before electroless plating, and the adhesion of the plated electrodes to the PC surface was strengthened with thermal annealing. Compared with the previously reported electroless plating technique for fabrication of microelectrodes on a microchip, the present method avoided the use of a membrane stencil with an electrode pattern to restrict the area to be wet-chemically sensitized. The CE with integrated ED (CE-ED) microchip was assembled by thermal bonding an electrode-plated PC cover plate to a microchannel-embossed PC substrate. The novel method allows one to fabricate low-cost, electrode-integrated, complete PC CE-ED chips with no need of a clean room. The fabricated CE-ED microchip was demonstrated for separation and detection of model analytes, including dopamine (DA) and catechol (CA). Detection limits of 0.65 and 1.03 microM were achieved for DA and CA, respectively, and theoretical plate number of 1.4 x 10(4) was obtained for DA. The plated gold electrode can be used for about 4 h, bearing usually more than 100 runs before complete failure.     

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.     

Underivatized cyclic olefin copolymer as substrate material and stationary phase for capillary and microchip electrochromatography

We report, for the first time, the use of underivatized cyclic olefin copolymer (COC, more specifically: Topas) as the substrate material and the stationary phase for capillary and microchip electrochromatography (CEC), and demonstrate chromatographic separations without the need of coating procedures. Electroosmotic mobility measurements in a 25 microm id Topas capillary showed a significant cathodic EOF that is pH-dependent. The magnitude of the electroosmotic mobility is comparable to that found in glass substrates and other polymeric materials. Open-tubular CEC was employed to baseline-separate three neutral compounds in an underivatized Topas capillary with plate heights ranging from 5.3 to 12.7 microm. The analytes were detected using UV absorbance at 254 nm, thus taking advantage of the optical transparency of Topas at short wavelengths. The fabrication of a Topas-based electrochromatography microchip by nanoimprint lithography is also presented. The microchip has an array of pillars in the separation column to increase the surface area. The smallest features that were successfully imprinted were around 2 microm wide and 5 microm high. No plasma treatment was used during the bonding, thus keeping the surface properties of the native material. An RP microchip electrochromatography separation of three fluorescently labeled amines is demonstrated on the underivatized microchip with plate heights ranging from 3.4 to 22 microm.     

Screen‐Printed Contactless Conductivity Detector for Microchip Capillary Electrophoresis

A new method for mass fabrication of silver ink conductivity detector electrodes for poly(methylmethacrylate) (PMMA) microchip electrophoretic systems has been developed based on screen‐printing technology. Printing of silver conductivity electrodes was performed through a patterned stencil on thin PMMA sheets. Following the electrode fabrication, the PMMA sheets are cut into cover sheets, and are aligned and sealed to the channel plate thus establishing a complete microchip separation device. The effects of the electrode width and spacing on the response and resolution have been investigated and the optimized electrode performance was compared to commonly used aluminum electrodes in the determination of ammonium, methyl ammonium, and sodium. The utility of the screen‐printed contactless conductivity detector (SPCCD) electrodes is further demonstrated for the separation and detection of organic acids with excellent reproducibility (RSD values of 3.7% and 4.1% for oxalate and tartrate, respectively). The thick‐film fabrication of the electrode material demonstrates the ability to mass‐fabricate detection devices with total process of device fabrication requiring less than 4 h (including the fabrication of channel plate, cover sheet with the electrodes, and subsequent bonding). The fabrication method described here is convenient and does not compromise the detector performance, hence offers great promise for producing single use field deployable analytical microsystems.     

Prototyping disposable electrophoresis microchips with electrochemical detection using rapid marker masking and laminar flow etching

Two novel methods are described for the fabrication of components for microchip capillary electrophoresis with electrochemical detection (microchip CEEC) on glass substrates. First, rapid marker masking is introduced as a completely nonphotolithographic method of patterning and fabricating integrated thin-film metal electrodes onto a glass substrate. The process involves applying the pattern directly onto the metal layer with a permanent marker that masks the ensuing chemical etch. The method is characterized, and the performance of the resulting electrode is evaluated using catecholamines. The response compares well with photolithographically defined electrodes and exhibits detection limits of 648 nM and 1.02 microM for dopamine and catechol, respectively. Second, laminar flow etching is introduced as a partially nonphotolithographic method of replicating channel networks onto glass substrates. The replication process involves applying a poly(dimethylsiloxane) (PDMS) mold of the channel network onto a slide coated with a sacrificial metal layer and then pulling solutions of metal etchants through the channels to transfer the pattern onto the sacrificial layer. The method is tested, and prototype channel networks are shown. These methods serve to overcome the time and cost involved in fabricating glass-based microchips, thereby making the goal of a disposable high performance lab-on-a-chip more attainable.     

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.     

In-situ grafting hydrophilic polymer on chitosan modified poly(dimethylsiloxane) microchip for separation of biomolecules

In this paper, a simple and green modification method is developed for biomolecules analysis on poly(dimethylsiloxane) (PDMS) microchip with successful depression of nonspecific biomolecules adsorption. O-[(N-succinimdyl)succiny]-o'-methyl-poly(ethylene glycol) was explored to form hydrophilic surface via in-situ grafting onto pre-coated chitosan (Chit) from aqueous solution in the PDMS microchannel. The polysaccharide chains backbone of Chit was strongly attracted onto the surface of PDMS via hydrophobic interaction combined with hydrogen bonding in an alkaline medium. The methyl-poly(ethylene glycol) (mPEG) could produce hydrophilic domains on the mPEG/aqueous interface, which generated brush-like coating in this way and revealed perfect resistance to nonspecific adsorption of biomolecules. This strategy could greatly improve separation efficiency and reproducibility of biomolecules. Amino acids and proteins could be efficiently separated and successfully detected on the coated microchip coupled with end-channel amperometric detection at a copper electrode. In addition, it offered an effective means for preparing biocompatible and hydrophilic surface on microfluidic devices, which may have potential use in the biological analysis.     

Fabrication of SU-8 based microchip electrophoresis with integrated electrochemical detection for neurotransmitters

A new SU-8 based microchip capillary electrophoresis (MCE) device has been developed for the first time with integrated electrochemical detection. Embedded electrophoretic microchannels have been fabricated with a multilayer technology based on bonding and releasing steps of stacked SU-8 films. This technology has allowed the monolithic integration in the device of the electrochemical detection system based on platinum electrodes. The fabrication of the chips presented in this work is totally compatible with reel-to-reel techniques, which guarantee a low cost and high reliability production. The influence of relevant experimental variables, such as the separation voltage and detection potential, has been studied on the SU-8 microchip with an attractive analytical performance. Thus, the effective electrical isolation of the end-channel amperometric detector has been also demonstrated. The good performance of the SU-8 device has been proven for separation and detection of the neurotransmitters, dopamine (DA) and epinephrine (EP). High efficiency (30,000-80,000 N/m), excellent precision, good detection limit (450 nM) and resolution (0.90-1.30) has been achieved on the SU-8 microchip. These SU-8 devices have shown a better performance than commercial Topas (thermoplastic olefin polymer of amorphous structure) microchips. The low cost and versatile SU-8 microchip with integrated platinum film electrochemical detector holds great promise for high-volume production of disposable microfluidic analytical devices.     

Chemical modification of polymeric microchip devices

Analytical polymeric microchips in both fluidic and array formats offer short analysis times, coupling of many sample processing and chemical reaction steps on one platform with minimal sample and reagent consumption, as well as low cost, minimal fabrication times and disposability. However, the invariable bulk properties of most commercial polymers have driven researchers to develop new modification strategies. This article critically reviews the scope and development of chemical modifications of such polymeric chips since 2003. Surface modifications were based on chemical derivatization or activation of surface layers with reagent solutions, reactive gases and irradiation. Bulk modification of polymer chips used newly incorporation of monomers with selective chemical functionalities throughout the bulk polymer material and integrated the chip modification and fabrication into a single step. Such modifications hold a great promise for establishing a true ‘lab-on-chip’ as can be seen from many novel applications for modulating electroosmosis, suppressing protein adsorption in microchip capillary electrophoretic separations, extraction of analytes and for zone-specific binding of enzymes and other biomolecules.     

Low temperature bonding of poly(methylmethacrylate) electrophoresis microchips by in situ polymerisation

A novel method for bonding poly(methyl methacrylate) (PMMA) electrophoresis microchips at the temperature below the glass transition temperature of PMMA based on in situ polymerization has been demonstrated. Methyl methacrylate (MMA) containing initiators was allowed to prepolymerize in an 85 degrees C water bath for 8 min and 15 min to produce a bonding solution and a dense molding solution, respectively. The channel plate of the PMMA microchip was fabricated by the UV-initiated polymerization of the molding solution between a nickel template and a PMMA plate at room temperature. Prior to bonding, the blank cover was coated with a thin layer of the bonding solution and was bonded to the channel plate at 95 degrees C for 20 min under the pressure of binder clips. The attractive performance of the PMMA chips bonded by the new approach has been demonstrated by separating and detecting dopamine, catechol, three cations, and three organic acids in connection with end-column amperometric detection and contactless conductivity detection.     

A disposable laser print-cut-laminate polyester microchip for multiplexed PCR via infra-red-mediated thermal control

Infrared (IR)-mediated thermal cycling system, a method proven to be a effective for sub-μL scale polymerase chain reaction (PCR) on microchips, has been integrated with DNA extraction and separation on a glass microchip in a fully integrated micro Total Analysis System by Easley et al., in 2006. IR-PCR has been demonstrated on both glass and PMMA microdevices where the fabrication (bonding) is not trivial. Polyester-toner (PeT) microfluidic devices have significant potential as cost-effective, disposable microdevices as a result of the ease of fabrication (∼$0.25 USD and <10 min per device) and availability of commercial substrates. For the first time, we demonstrate here the thermal cycling in PeT microchips on the IR-PCR system. Undesirable IR absorption by the black-toner bonding layer was eliminated with a spatial filter in the form of an aluminum foil mask. The solution heating rate for a black PeT microchip using a tungsten lamp was 10.1 ± 0.7 °C s⁻¹ with a cooling rate of roughly −12 ± 0.9 °C s⁻¹ assisted by forced air cooling. Dynamic surface passivation strategies allowed the successful amplification of a 520 bp fragment of the λ-phage genome (in 11 min) and a 1500 bp region of Azospirillum brasilense. Using a centrosymmetric chamber configuration in a multichamber PeT microchip, homogenous temperature distribution over all chambers was achieved with inter-chamber temperature differences at annealing, extension and denaturing steps of less than ±2 °C. The effectiveness of the multichamber system was demonstrated with the simultaneous amplification of a 390 bp amplicon of human β-globin gene in five PeT PCR microchambers. The relative PCR amplification efficiency with a human β-globin DNA fragment ranged from 70% to 90%, in comparison to conventional thermal cyclers, with an inter-chamber standard deviation of ∼10%. Development of PeT microchips for IR-PCR has the potential to provide rapid, low-volume amplification while also integrating PCR with extraction upstream and separation/detection downstream.     

Microchip PCR

Miniaturization of genetic tests has become an important goal. This review surveys the current progress towards the miniaturization of tests based on the polymerase chain reaction (PCR). It examines the different types of PCR microchip designs, fabrication methods,and the components of a microchip PCR device. It also discusses the problems attributable to surface chemistry of microchip components (inhibition of PCR), and the static and dynamic surface passivation strategies developed for the solution of these difficulties     

A sol-gel-modified poly(methyl methacrylate) electrophoresis microchip with a hydrophilic channel wall

A sol-gel method was employed to fabricate a poly(methyl methacrylate) (PMMA) electrophoresis microchip that contains a hydrophilic channel wall. To fabricate such a device, tetraethoxysilane (TEOS) was injected into the PMMA channel and was allowed to diffuse into the surface layer for 24 h. After removing the excess TEOS, the channel was filled with an acidic solution for 3 h. Subsequently, the channel was flushed with water and was pretreated in an oven to obtain a sol-gel-modified PMMA microchip. The water contact angle for the sol-gel-modified PMMA was approximately 27.4 degrees compared with approximately 66.3 degrees for the pure PMMA. In addition, the electro-osmotic flow increased from 2.13x10(-4) cm2 V(-1) s(-1) for the native-PMMA channel to 4.86x10(-4) cm2 V(-1) s(-1) for the modified one. The analytical performance of the sol-gel-modified PMMA microchip was demonstrated for the electrophoretic separation of several purines, coupled with amperometric detection. The separation efficiency of uric acid increased to 74,882.3 m(-1) compared with 14,730.5 m(-1) for native-PMMA microchips. The result of this simple modification is a significant improvement in the performance of PMMA for microchip electrophoresis and microfluidic applications.     

Atmospheric molded poly(methylmethacrylate) microchip emitters for sheathless electrospray

Disposable poly(methylmethacrylate) (PMMA) sheathless electrospray microchip emitters were prepared for the first time using the atmospheric molding fabrication protocol. A sheathless electrospray from uncoated channel outlets, machined to cone-shaped three-dimensional tips, is demonstrated utilizing a simple cross design with an on-chip liquid junction to obviate the need for external unions to voltage electrodes, thus reducing the dead volume effects as well as the complexity of fabrication. The fast replication of microchip emitters was performed by molding prepolymeric methylmethacrylate solutions into silicon-master/aluminum-spacer/glass-plate molds followed by UV-initiated free radical polymerization. The performance of the new microchip emitters was demonstrated for mass spectral measurements of methionine enkephalin, adrenocorticotropic hormone and insulin peptide/protein mixtures. The samples were infused through capillary connections using hydrodynamic pumping. The polymeric emitters prepared by this flexible fabrication route offer an easy way of operation and high stability, without a need for attachment of external voltage unions or metallizing the emitter tips. The new approach should provide a useful low-cost tool for widespread coupling of mass spectrometry to chip systems.     

A rejuvenation method for poly(N,N-dimethylacrylamide)-coated glass microfluidic chips

As microfluidic chips come to integrate the higher levels of functionality required for the implementation of advanced bioanalytical protocols, a crucial factor is that of cost. Although glass chips provide advantages in multilayer integrations, their cost is far higher than that of polymer chips. However, a simple and effective rejuvenation protocol for glass microchips may enable higher levels of integration and functionality on glass microchips. Here we present a method to rejuvenate glass microchips that had been used for capillary electrophoresis to the extent that their performance was degraded. This degradation was due to one of the two mechanisms: (i) a deterioration of the polymer coating on the inner surface of the microchannel or (ii) an aging of the glass substrate. Using the method presented here, we have rejuvenated more than 50 such "aged" microchips. The performance of these microchips was fully restored after the rejuvenation and lasted for hundreds of DNA separation runs. Our experiments indicate that the loss of resolution in microchip separations was not associated with glass aging, but was due to the degradation of the polymer coating on the inner surface of microchannels. This suggests that it is possible to extend the microchip lifetime "forever" using the rejuvenation protocol and that the exploration of higher levels of integration and functionality on glass microchips (or of hybrid structures involving materials capable of withstanding the reagents and elevated temperatures used) is feasible.     

Fracture mechanism of metal electrode integrated on a chip and fabrication of a poly(ethylene terephthalate) electrophoresis microchip

Thermal bonding is an important technique to fabricate polymer electrophoresis microchip. However, the metal electrodes deposited on polymer substrate can readily fracture during the thermal bonding. In this paper, poly(ethylene terephthalate) (PET) was exploited to fabricate the electrophoresis microchip with an integrated gold electrode for amperometric detection. The fracture of the gold electrode was studied through FEA (finite element analysis) simulations, the potentially risk positions on the electrode were shown. The calculation results were tested by bonding experiments and were proven to be consistent with the experiments. Besides, an optimal bonding temperature for PET chip was also presented based on FEA simulations and bonding experiments. Considering the low surface properties of PET, oxygen plasma-assisted thermal bonding technique was used to enhance bonding. Upon treated for 150 s, the PET substrates could be thermally bonded at 62 °C without electrode fracture. The fabricated PET chips were demonstrated for detection of standard glucose solution. Satisfactory reproducibility was achieved, and the RSD values of peak height and migration time of the PET CE chips were 0.51% and 2.17%, respectively.     

Rapid bonding of Pyrex glass microchips

A newly developed vacuum hot press system has been specially designed for the thermal bonding of glass substrates in the fabrication process of Pyrex glass microchemical chips. This system includes a vacuum chamber equipped with a high-pressure piston cylinder and carbon plate heaters. A temperature of up to 900 degrees C and a force of as much as 9800 N could be applied to the substrates in a vacuum atmosphere. The Pyrex substrates bonded with this system under different temperatures, pressures, and heating times were evaluated by tensile strength tests, by measurements of thickness, and by observations of the cross-sectional shapes of the microchannels. The optimal bonding conditions of the Pyrex glass substrates were 570 degrees C for 10 min under 4.7 N/mm(2) of applied pressure. Whereas more than 16 h is required for thermal bonding with a conventional furnace, the new system could complete the whole bonding processes within just 79 min, including heating and cooling periods. Such improvements should considerably enhance the production rate of Pyrex glass microchemical chips. Whereas flat and dust-free surfaces are required for conventional thermal bonding, especially without long and repeated heating periods, our hot press system could press a fine dust into glass substrates so that even the areas around the dust were bonded. Using this capability, we were able to successfully integrate Pt/Ti thin film electrodes into a Pyrex glass microchip.     

Magnetron sputtering Si interlayer: a protocol to prepare solid phase microextraction coatings on metal-based fiber

Use of metal fibers in solid phase microextraction (SPME) can overcome the fragility drawback of conventional fused-silica ones. However, the surface modification of metal substrates is rather difficult, which largely prevents many mature traditional techniques, such as sol-gel and chemical bonding, being used in fabrication of SPME coating on metal-based fibers. This study demonstrates a protocol to resolve this problem by magnetron sputtering a firm Si interlayer on stainless steel fiber. The Si interlayer was easily modified active group, and attached with a multiwalled carbon nanotubes (MWCNTs) coating using the reported approach. The as-prepared MWCNTs/Si/stainless steel wire fiber not only preserved the excellent SPME behaviors of MWCNTs coatings, but also exhibited a number of advantages including high rigidity, long service life, and good stability at high temperature, in acid and alkali solutions. This new surface modification technique might provide a versatile approach to prepare sorbent coatings on unconfined substrates using traditional methods.