Characterization of a new fluorescence-enhancing substrate for microarrays with femtomolar sensitivity

The demand for high sensitivity in microarray technology has stimulated the research of new labeling strategies, new substrates with increased loading capacity and new approaches to amplify the fluorescence signals. Although these methods improve the sensitivity, they are based on non-routine procedures which limit their widespread usage. A simple way to achieve fluorescence enhancement is the optical interference (OI) coating technology based on the use of substrates with films of well-defined thickness that maximize photoabsorption of the dye molecules in the vicinity of the surface and reflect the emitted light toward the detector. Here we introduce a new substrate for fluorescence enhancement composed of a thin aluminum mirror and a single quarter wave silicon oxide as a dielectric layer. The use of such material allows the development of a substrate suited for parallel production in conventional IC (Integrated Circuit's) technology, which produces a 20-fold enhancement of fluorescence compared to glass, which is the most common material used in microarrays. The new substrate was coated with a copolymer of N,N-dimethylacrylamide, N-acryloyloxysuccinimide, and 3-(trimethoxysilyl)propyl methacrylate, copoly(DMA-NAS-MAPS), which forms, by a simple and robust procedure, a functional nanometric film that covalently binds bio-probes on the surface and efficiently suppresses non-specific adsorption. The performance of the new fluorescence-enhancing substrate in microarray technology was demonstrated for the detection of a panel of inflammation biomarkers pushing detection limits into the femtomolar range. (C) 2013 Elsevier B.V. All rights reserved.