Fabrication of nanofluidic biochips with nanochannels for applications in DNA analysis

With the development of nanotechnology, great progress has been made in the fabrication of nanochannels. Nanofluidic biochips based on nanochannel structures allow biomolecule transport, bioseparation, and biodetection. The domain applications of nanofluidic biochips with nanochannels are DNA stretching and separation. In this Review, the general fabrication methods for nanochannel structures and their applications in DNA analysis are discussed. These representative fabrication approaches include conventional photolithography, interference lithography, electron-beam lithography, nanoimprint lithography and polymer nanochannels. Other nanofabrication methods used to fabricate unique nanochannels, including sub-10-nm nanochannels, single nanochannels, and vertical nanochannels, are also mentioned. These nanofabrication methods provide an effective way to form nanoscale channel structures for nanofluidics and biosensor devices for DNA separation, detection, and sensing. The broad applications of nanochannels and future perspectives are also discussed.     

Soft lithographic printing of patterns of stretched DNA and DNA/electronic polymer wires by surface-energy modification and transfer

Aligned and stretched lambda DNA is directed to specific locations on solid substrates. Surface-energy modification of glass substrates by using patterned polydimethylsiloxane (PDMS) stamps is used to direct DNA onto the surface-energy-modified micrometer-scale pattern through molecular combing. As an alternative, patterned and nonpatterned PDMS stamps modified with polymethylmethacrylate (PMMA) are utilized to direct the stretched DNA to the desired location and the results are compared. The DNA is elongated through molecular combing on the stamp and transfer printed onto the surfaces. PMMA-modified stamps show a more defined length of the stretched DNA, as compared to bare PDMS stamps. A combination of these two methods is also demonstrated. As an application example, transfer printing of DNA decorated with a semiconducting conjugated polyelectrolyte is shown. The resulting patterned localization of stretched DNA can be utilized for functional nanodevice structures, as well as for biological applications.     

Transparent conductive thin film synthesis based on single-walled carbon nanotubes dispersion containing polymethylmethacrylate binder

Transparent conductive hybrid thin films of single-walled carbon nanotubes (SWNTs) and polymethyl methacrylate (PMMA) are fabricated using dispersions containing SWNTs and water-borne PMMA binder. The polymer binder was used as adhesion promoter between the SWNTs and the substrate. The polymer binder content in the SWNTs dispersion is varied to obtain the optimum optical transmittance, electrical conductivity, and mechanical adhesion. The PMMA and SWNT network formed the composite over substrate. The fabricated SWNTs/PMMA hybrid films are immersed in nitric acid (HNO3) and thionyl chloride (SOCl2) to improve electrical conductivity. SWNTs films with 0.2-0.6 mg/ml polymer binder have sheet resistance of 80-140 ohms/sq at a transmittance of about 80% and a strong adhesion on glass substrate. Furthermore, the electrical stability of the films is improved via the PMMA addition. This results indicates that the SWNTs/PMMA hybrid films fabricated by this method can be used as an alternative of indium tin oxide (ITO) film on flexible substrate.     

Control of Polymer Phase Separation by Roughness Transfer Printing for 2D Microlens Arrays

Great efforts have been devoted to the control of phase separation between blended polymers in terms of the advantages for engineering functional topologies. A simple and straightforward pathway through roughness transfer printing (RTP) is proposed to realize the control of polymer phase separation. The additional roughness difference, which is introduced by trace agarose transferred from a hydrogel stamp, offers a great effect on the rate of nucleation and coalescence orientation of polymethylmethacrylate (PMMA) protrusions grown from a polydimethylsiloxane (PDMS) network. Using a particular topography of agarose stamp and a proper growth time in toluene atmosphere, a 2D microlens array with high uniformity is obtained that shows great potential for optical applications. Moreover, the control of polymer phase separation was successfully extended to the collection and identification of fingerprints with a high degree of replication.     

Soft lithography meets self-organization: Some new developments in meso-patterning

This is a brief review of our recent and ongoing work on simple, rapid, room temperature, pressure-less and large area (∼ cm²) imprinting techniques for high fidelity meso-patterning of different types of polymer films. Examples include soft solid polymer films and surfaces like cross-linked polydimethylsiloxane (PDMS) and polyacrylamide (PAA) based hydrogels, thermoplastics like polystyrene (PS), polymethylmethacrylate (PMMA) etc both on planar and curved surfaces. These techniques address two key issues in imprinting: (i) attainment of large area conformal contact with the stamp, especially on curved surfaces, and (ii) ease of stamp detachment without damage to the imprinted structures. The key element of the method is the use of thin and flexible patterned foils that readily and rapidly come into complete conformal contact with soft polymer surfaces because of adhesive interfacial interactions. The conformal contact is established at all length scales by bending of the foil at scales larger than the feature size, in conjunction with the spontaneous deformations of the film surface on the scale of the features. Complex two-dimensional patterns could also be formed even by using a simple one-dimensional master by multiple imprinting. The technique can be particularly useful for the bulk nano applications requiring routine fabrication of templates, for example, in the study of confined chemistry phenomena, nanofluidics, bio-MEMS, micro-imprinting, optical coatings and controlled dewetting.     

Finite element method simulation of the molding process for thermal nano-imprint lithography

We made a numerical study on the deformation of a viscoelastic polymethyl methacrylene (PMMA) resist when a rigid SiO2 stamp with a rectangular line pattern is imprinted into the PMMA resist for thermal nano-imprint lithography (NIL). The stress distribution in the polymer resist during the molding process is calculated by a finite element method (FEM). Our simulation results reveal that the asymmetric von Mises stress is distributed over the polymer around the external line, which seems to be due to the squeezing flow under the flat space. The stress seems to be concentrated at the sidewall close to the centerline of the whole structure. Our simulation also reveals that a micro gap is formed between the replicated structure and the outer wall of the mold.     

Entropic unfolding of DNA molecules in nanofluidic channels

Single DNA molecules confined to nanoscale fluidic channels extend along the channel axis in order to minimize their conformational free energy. When such molecules are forced into a nanoscale fluidic channel under the application of an external electric field, monomers near the middle of the DNA molecule may enter first, resulting in a folded configuration with less entropy than an unfolded molecule. The increased free energy of a folded molecule results in two effects: an increase in extension factor per unit length for each segment of the molecule, and a spatially localized force that causes the molecule to spontaneously unfold. The ratio of this unfolding force to hydrodynamic friction per DNA contour length is measured in nanochannels with two different diameters.     

Stamp design effect on 100 nm feature size for 8 inch NanoImprint lithography

Sub-100?nm resolution on a 200?mm silicon stamp has been hot embossed into commercial Sumitomo NEB 22 resist. A single pattern, exposed with electron beam lithography, has been considered to define the stamp and thus make it possible to point out the impact of stamp design on the printing. These results may be considered as a first attempt to define rules to solve the proximity printing effects (PPEs). Moreover, a large range of initial resist thickness, from 56 to 506?nm, has been spin coated to assess the effect of polymer flow properties for the stamp cavity filling and the printed defects. A?detailed analysis of the printed resist in dense hole patterns showed that the application volume conservation is enough to calculate the residual layer thickness as the height of the printed resist feature. Good accordance has been obtained between the theoretical approach and experimental results. Moreover, the impact of the pattern symmetry breakdown on mould deformation is clearly shown in this paper in the printed areas as well as in the unprinted areas.     

Design and fabrication of label-free biochip using a guided mode resonance filter with nano grating structures by injection molding process

This paper introduces technology to fabricate a guided mode resonance filter biochip using injection molding. Of the various nanofabrication processes that exist, injection molding is the most suitable for the mass production of polymer nanostructures. Fabrication of a nanograting pattern for guided mode resonance filters by injection molding requires a durable metal stamp, because of the high injection temperature and pressure. Careful consideration of the optimized process parameters is also required to achieve uniform sub-wavelength gratings with high fidelity. In this study, a metallic nanostructure pattern to be used as the stamp for the injection molding process was fabricated using electron beam lithography, a UV nanoimprinting process, and an electroforming process. A one-dimensional nanograting substrate was replicated by injection molding, during which the process parameters were controlled. To evaluate the geometric quality of the injection molded nanograting patterns, the surface profile of the fabricated nanograting for different processing conditions was analyzed using an atomic force microscope and a scanning electron microscope. Finally, to demonstrate the feasibility of the proposed process for fabricating guided mode resonance filter biochips, a high-refractive-index material was deposited on the polymer nanograting and its guided mode resonance characteristics were analyzed.     

Fabrication of fluorine-doped diamond-like carbon stamps for UV nanoimprint lithography

Two fluorine-doped diamond-like carbon (F-DLC) stamps with sub-100 nm line patterns were fabricated using a direct etching method. These were applied successfully to ultraviolet (UV) nanoimprint lithography without requiring an anti-adhesion layer coating. Tests were performed to determine the optimum fluorine concentration for the F-DLC stamps. The ideal stamp material consisted of 25?at.% F-DLC with a contact angle of 85°, UV transmittance of 16.4-26.8%, and hardness of 4.5?GPa. The O(2) plasma etch rate of the DLC was increased considerably by the fluorine doping, making it comparable to the etch rate of polymethyl methacrylate (PMMA). Thus, only PMMA was used as the etch mask in the fabrication of the stamps.     

Thermally assisted semiconductor-like to insulator transition in gold-poly(methyl methacrylate) nanocomposites

Gold-polymethylmethacrylate (PMMA) nanocomposites were fabricated by mixing gold nanoparticles capped with oleylamine in polymethylmethacrylate. The samples were analysed using UV-vis absorption spectroscopy, transmission electron microscopy, small angle x-ray scattering, Fourier transform infrared spectrometry (FTIR) and x-ray photoelectron spectroscopy (XPS). Electrical resistivity of nanocomposite samples was measured by a four-probe technique in the 70-300?K range. The nanocomposites showed a transition with an onset at ～160-165?K. They exhibited a semiconductor-like conductivity at higher temperatures and nearly temperature independent conductivity at lower temperatures. The interfacial interaction of Au nanoparticles and PMMA polymer is investigated using FTIR and XPS. A ligand-exchange process occurs when capped gold nanoparticles are incorporated in PMMA polymer.     

DNA-functionalized nanochannels for SNP detection

We have developed ultrahigh density array of functionalized nanochannels by using a block copolymer having end di-COOH group. This approach provides a facile route for direct functionalization of wall surface of the nanochannels and immobilization site for molecular recognition agents (MRAs). By using overhanging single-stranded DNA as MRAs, the DNA-functionalized nanochannels showed high resolution to detect a single-base mismatch as well as to discriminate single-mismatched sequence at various locations by hybridization preference with MRAs.     

Parallel-local anodic oxidation of silicon surfaces by soft stamps

We investigate the fabrication of nanometric patterns on silicon surfaces by using the parallel-local anodic oxidation technique with soft stamps. This method yields silicon oxide nanostructures 15?nm high, namely at least five times higher than the nanostructures made with local anodic oxidation using atomic force microscopy, and thanks to the size of the stamp enables one to pattern the surface across a centimetre length scale. To implement this technique, we built a machine to bring the metallized polydimethylsiloxane stamp in contact with the silicon surface, subsequently inserted in a sealed chamber with controlled relative humidity. The oxide nanostructures are fabricated when a bias voltage of 36?V is applied between the stamp and the silicon for 2?min, with a relative humidity of 90%. The flexibility of the stamp enables a homogeneous conformal contact with the silicon surface, resulting in an excellent reproducibility of the process. Moreover, by means of two subsequent oxidations with the same stamp and just rotating the sample, we are able to fabricate complex nanostructures. Finally, a detailed study of the oxidation mechanism, also using a finite element analysis, has been performed to understand the underlying mechanism.     

Synthesis and characterization of SiO2/(PMMA/Fe3O4) magnetic nanocomposites

Magnetic silica nanocomposites (magnetic nanoparticles core coated by silica shell) have the wide promising applications in the biomedical field and usually been prepared based on the famous St?ber process. However, the flocculation of Fe3O4 nanoparticles easily occurs during the silica coating, which limits the amount of magnetic silica particles produced in the St?ber process. In this paper, PMMA/Fe3O4 nanoparticles were used in the St?ber process instead of the "nude" Fe3O4 nanoparticles. And coating Fe3O4 with PMMA polymer beforehand can prevent magnetic nanoparticles from the aggregation that usually comes from the increasing of ionic strength during the hydrolyzation of tetraethoxysilane (TEOS) by the steric hindrance. The results show that the critical concentration of magnetic nanoparticles can increase from 12 mg/L for "nude" Fe3O4 nanoparticles to 3 g/L for PMMA/Fe3O4 nanoparticles during the St?ber process. And before the deposition of silica shell, the surface of PMMA/FeO4 nanoparticles had to be further modified by hydrolyzing them in CH3OH/NH3 x H2O mixture solution, which provides the carboxyl groups on their surface to react further with the silanol groups of silicic acid.     

Hybrid dynamic coating with n-dodecyl beta-D-maltoside and methyl cellulose for high-performance carbohydrate analysis on poly(methyl methacrylate) chips

Hybrid dynamic coating using n-dodecyl beta-d-maltoside (DDM) and methyl cellulose (MC) has been developed for suppression of analyte adsorption and electroosmotic flow (EOF) in a poly(methyl methacrylate) (PMMA) channel. The adsorption of APTS-labeled sugars in a PMMA channel was obviously suppressed with DDM dynamic coating; however, EOF was reduced only by a factor of approximately 25%, resulting in irreproducible separations. In contrast, both analyte adsorption and EOF in a PMMA channel were efficiently minimized with MC coating; however, concentrated MC above 0.3% was required to achieve high-performance separations, which greatly increased viscosity of the solution and caused difficulties during buffer loading and rinsing. In addition, n-dodecyltrimethylammonium chloride did not show observable effects on reducing analyte adsorption, although it has the same hydrophobic alkyl chain as DDM. These results strongly indicated that the polysaccharide moiety of surface modifiers has a specific affinity to surface charges and is crucial to achieving efficient and stable dynamic coating on the PMMA surface. Hybrid dynamic coating with 0.25% DDM and 0.03% MC was found to minimize both analyte adsorption and EOF in a PMMA channel to a negligible level, while still keeping a low viscosity of the solution. High-speed and high-throughput profiling of the N-linked glycans derived from alpha1-acid glycoprotein, fetuin, and ribonuclease B was demonstrated in both single-channel and 10-channel PMMA chips using DDM-MC hybrid coating. We propose that DDM-MC hybrid coating might be a general method for suppressing analyte adsorption and EOF in polymer MCE devices. The current MCE-based method might be a promising alternative for high-throughput screening of carbohydrate alterations in glycoproteins.     

Near-white light luminescent material from Eu(III) complexes encapsulated in silica/PMMA matrices

A novel ternary molecular hybrid material has been obtained by immobilization of Eu-Salen complex into silica matrix and poly (methyl methacrylate) (PMMA) matrix. Here, the Salen-type Schiff-base ligand H₂Salen (N,N′-bis(salicylidene)ethylenediamine) which has been successfully modified by 3-(triethoxysilyl)-propyl isocyanate (TESPIC) has been used as a flexible linker and the antenna. The obtained solid hybrid material shows not only the characteristic red emission of Eu³⁺ but also the blue emission of Salen-Si host arising from the inefficient energy transfer from antenna to Eu³⁺, leading to the unexpected near-white light color of the material. For comparison, the binary hybrid without PMMA has also been prepared. Photoluminescent spectra suggest that the introduction of PMMA can enhance the emission intensity whereas the chromaticity nearly has not been changed. Moreover, thermal analysis has revealed the good thermal stability of the ternary hybrid material. X-ray diffraction patterns demonstrate the amorphous structure of both hybrid materials.     

Thermal expansion and contraction of an elastomer stamp causes position-dependent polymer patterns in capillary force lithography

It is often observed that polymer patterns fabricated by capillary force lithography (CFL) are not identical, position-dependent even in one sample. The drawback has not been successfully explained so far. This paper reveals that the position-dependent pattern is mainly caused by the volume expansion and contraction of the elastomer stamp during heating and cooling in the CFL process. The stamp expands on a polymer liquid on heating, accumulating the polymer at one side-wall of each pattern of the stamp. And the stamp shrinks back to the initial position, accumulating the polymer at the opposite wall of the stamp pattern. For crystalline polymers, the morphology was mainly determined by the annealing temperature, that is, the degree of expansion. The position-dependence of the morphology was enhanced as the annealing temperature was increased. For amorphous polymers, the morphology was sensitive to cooling rate. Fast cooling led to a frozen morphology generated at the hot annealing temperature, while slow cooling produced an opposite morphology from the one at the annealing. The experimental results were theoretically explained by analyzing thermal expansion of the stamp and the shear stress exerted in the polymer layer. In the conclusion, we added our suggestions to avoid the nonuniformity in the polymer pattern by CFL process.     

Low cost fabrication of the large-area anti-reflection films from polymer by nanoimprint/hot-embossing technology

In this paper, we present a low cost approach to produce large-area polymer sheets with sub-wavelength nanostructures. The fabricated polymer films would have great potentials to attach to optical or solar-cell-related consumer products when anti-reflection/anti-glaring is mandatory. We employed a special electron cyclotron resonance (ECR) plasma process to fabricate the SWSs with large area directly on silicon substrates. Homogeneously distributed nanotips on the full 4 inch silicon substrate were fabricated by using gas mixtures of silane, methane, argon and hydrogen. An Ni-Co metal mold with a hardness of 550 HV was produced through the replication of the Si mold by electroplating. Afterwards, the molding process was applied to manufacture the nanostructures in PMMA plates in large volume. The nanostructures in PMMA plates with aspect ratios of 4 and diameters of 150?nm were fabricated. The fabricated PMMA sheets could generate the gradient of the refractive indices, absorb the light and greatly reduce the reflectivity. Compared with the PMMA without SWSs, the reflectivity of PMMA with SWSs decreased dramatically from 4.25% to 0.5% at the wavelength of light from 400 to 800?nm.     

Study of poly(methyl methacrylate)/poly(ethylene oxide) blends in solution. I

Determination of polymer-polymer interaction parameters (b₂₃) has been made by viscometry for poly(methyl methacrylate)/poly (ethylene oxide) blends, in solution, of a wide range of molecular weights of both polymers. The results obtained show that the compatibility of the polymer pair PMMA/PEO in 1.2-dichloroethane depends not only on the molecular weight of the samples used but also on the concentration of one polymer in relation to another.     

PMMA brush-containing two-solution bone cement: preparation, characterization, and influence of composition on cement properties

Two-solution bone cement consisting of poly (methyl methacrylate) (PMMA) brushes in methyl methacrylate has been developed as an alternative to the traditional two-solution (TSBC) and powder-liquid cements. It was hypothesized that the substitution of brushes, for the entire pre-polymer phase of the cement, would permit a decrease in solution viscosity at higher polymer fractions, and allow for physical entanglements with the cement matrix. Consequently, improved cement exothermal and mechanical properties could be expected with brush addition. PMMA brushes were grafted on the surface of cross-linked PMMA nanospheres following a multi-stage synthetic strategy. Brushes exhibiting optimal molecular weight for preparation of TSBC were used for characterization of cement viscosity, flexural and compressive mechanical properties, exothermal properties and residual monomer content. Interactions between grafts and free polymer formed during free radical polymerization of the cement were evaluated based on molecular weight measurements of the cement matrix and brushes. Brush-containing cements exhibited lower viscosity at significantly higher polymer fractions in comparison to TSBC. Cements with PMMA brushes had significantly lower polymerization temperatures and residual monomer content. Measurements of molecular weight revealed the existence of a dry brush regime when using the brush compositions selected in this study, which led to a reduction in the mechanical properties of some of the compositions tested. The optimal cement viscosity and maintenance of other important cement properties achieved with addition of PMMA brushes is expected to expand the use of the two-solution cements in a range of applications.