Fluorescence Signal Amplification on the Gel Biochips with a Mirror Surface and Optimization of Immunoassay Procedure

摘要

Biological microchips-the matrices of M × N elements containing biological probes (DNA, RNA, oligo-nucleotides, proteins, peptides, oligosaccharides, pads, etc.)-make it possible to perform parallel multiparam-eter analysis of samples [1-8]. The probes are immobilized either on the surface of a substrate (2D), such as glass, plastic, and metal [1-4], or into the volume of three-dimensional gel pads (3D) fixed on a substrate (Figs, la, lb) [5-8]. It has been earlier demonstrated that use of metal-coated substrates when manufacturing surface microchips can influence the fluorescence intensity during analysis; moreover, the fluorescence intensity can both increase and decrease depending on several factors, such as the distance between fluoro-phore and metal surface and excitation conditions [9-13]. The goal of this work was to study the effect of fluorescence signal amplification for 3D hydrogel microchips immobilized on the substrate with a reflecting metal coating and estimate the efficiency of such microchips in a quantitative immunofluorescence assay.