RESUMO
A number of materials used to fabricate disposable microfluidic devices are hydrophobic in nature with water contact angles on their surface ranging from 80° to over 100°. This characteristic makes them unsuitable for a number of microfluidic applications. Both the wettability and analyte adsorption parameters are highly dependent on the surface hydrophobicity. In this article, we propose a general method to coat the surface of five materials: polydimethylsiloxane (PDMS), cyclic olefin copolymer (COC), polyethylene terephthalate (PET), polycarbonate (PC), and polytetrafluoroethylene (PTFE). This fast and robust process, which is easily implementable in any laboratory including microfabrication clean room facilities, was devised by combining gas-phase and wet chemical modification processes. Two different coatings that improve the surface hydrophilicity were prepared via the "dip and rinse" approach by immersing the plasma oxidized materials into an aqueous solution of two different poly(dimethylacrylamide) copolymers incorporating a silane moiety and functionalized with either N-acryloyloxysuccinimide (NAS) (poly(DMA-NAS-MAPS) or glycidyl methacrylate (GMA) (poly(DMA-GMA-MAPS). The coating formation was confirmed by contact angle (CA) analysis comparing the variation of CAs of uncoated and coated surfaces subjected to different aging treatments. The antifouling character of the polymer was demonstrated by fluorescence and interferometric detection of proteins adsorbed on the surafce. This method is of great interest in microfluidics due to its broad applicability to a number of materials with varying chemical compositions.
Assuntos
Interações Hidrofóbicas e Hidrofílicas , Técnicas Analíticas Microfluídicas/instrumentação , Polímeros/química , Adsorção , Técnicas de Cultura de Células , Dimetilpolisiloxanos/química , Microfluídica , Cimento de Policarboxilato/química , Polietilenoglicóis/química , Polietilenotereftalatos , Água/química , MolhabilidadeRESUMO
The interaction of carbohydrates with a variety of biological targets, including antibodies, proteins, viruses, and cells are of utmost importance in many aspects of biology. Glycan microarrays are increasingly used to determine the binding specificity of glycan-binding proteins. In this study, a novel microarray support is reported for the fabrication of glycan arrays that combines the higher sensitivity of a layered Si-SiO2 surface with a novel polymeric coating easily modifiable by subsequent click reaction. The alkyne-containing copolymer, adsorbed from an aqueous solution, produces a coating by a single step procedure and serves as a soft, tridimensional support for the oriented immobilization of carbohydrates via azide/alkyne Cu (I) catalyzed "click" reaction. The advantages of a functional 3D polymer coating making use of a click chemistry immobilization are combined with the high fluorescence sensitivity and superior signal-to-noise ratio of a Si-SiO2 substrate. The proposed approach enables the attachment of complex sugars on a silicon oxide surface by a method that does not require skilled personnel and chemistry laboratories.
Assuntos
Química Click/métodos , Análise em Microsséries/métodos , Polímeros/química , Polissacarídeos/química , Fluorescência , Polímeros/síntese química , Silício/químicaRESUMO
A new methodology for the fabrication of an high-performance peptide microarray is reported, combining the higher sensitivity of a layered Si-SiO2 substrate with the oriented immobilization of peptides using a N,N-dimethylacrylamide-based polymeric coating that contains alkyne monomers as functional groups. This clickable polymer allows the oriented attachment of azido-modified peptides via a copper-mediated azide/alkyne cycloaddition. A similar coating that does not contain the alkyne functionality has been used as comparison, to demonstrate the importance of a proper orientation for facilitating the probe recognition and interaction with the target antibody.
Assuntos
Proteínas Imobilizadas/química , Peptídeos/química , Polímeros/química , Adsorção , Alcinos/química , Azidas/química , Química Click , Análise Serial de Proteínas , Silanos/química , Silício/química , Dióxido de Silício/química , Propriedades de SuperfícieRESUMO
The recent technological advances in micro/nanotechnology present new opportunities to combine microfluidics with microarray technology for the development of small, sensitive, single-use, point-of-care molecular diagnostic devices. As such, the integration of microarray and plastic microfluidic systems is an attractive low-cost alternative to glass based microarray systems. This paper presents the integration of a DNA microarray and an all-polymer microfluidic foil system with integrated thin film heaters, which demonstrate DNA analysis based on melting curve analysis (MCA). A novel micro-heater concept using semi-transparent copper heaters manufactured by roll-to-roll and lift-off on polyethylene naphthalate (PEN) foil has been developed. Using a mesh structure, heater surfaces have been realized in only one single metallization step, providing more efficient and homogenous heating characteristics than conventional meander heaters. A robust DNA microarray spotting protocol was adapted on Parylene C coated heater-foils, using co-polymer poly(DMA-NAS-MAPS) to enable covalent immobilization of DNA. The heaters were integrated in a microfluidic channel using lamination foils and MCA of the spotted DNA duplexes showed single based discrimination of mismatched over matched target DNA-probes. Finally, as a proof of principle, we perform MCA on PCR products to detect the Leu7Pro polymorphism of the neutropeptide Y related to increased risk of Type II diabetes, BMI and depression.