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Abstract

A method and device, based on a film of a luminescent substance, such as colloidal semiconductor nanocrystals dispersed in a polymer matrix, for conducting quantitative and real-time analyses of PCR processes or of biomolecular interactions in genomics and/or proteomics. The optical detection system is based on FRET processes between the luminescent substance (which acts as the donor in the FRET process) and a suitable fluorophore (which acts as the acceptor species) with which the DNA or other biomolecule is marked. The device is essentially composed of a reaction microchamber with a wall formed by a thin film made of polymer material, in which the nanocrystals are uniformly dispersed, or made of a photoluminescent or electroluminescent polymer. Molecular probes are chemically immobilized on the surface of the polymer film for the specific recognition of the analyte which is to be determined in real time. The film of nanocrystals is excited by radiation at low wavelength (for example, UV/blue), and the radiation in the spectral emission window characteristic of the fluorescent marker of the biomolecule is detected. The specific photophysical characteristics of FRET processes make it possible to monitor in a selective way, in real time and in quantitative mode, the biomolecular interactions, which take place in the close proximity of the surface of the film (typically at distances of <10 nanometres), thus almost completely reducing possible interference caused by background signals and by the free biomolecules in solution (which have not interacted with the corresponding recognition sites). The characteristics of the device also enable simultaneous analyses to be conducted in parallel on different biomolecules (multiplexing).

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