Single molecule detection using charge-coupled device array technology. Final report

摘要

An ultra sensitive technique for the detection of fluorescent species in a flowing stream has been developed. The extension of this technique to the detection of fluorescently tagged nucleotides will be a significant benefit to one of the novel approaches for DNA sequencing being developed at Los Alamos National Laboratories. The detection scheme is based on a novel mode of operating a Charge-Coupled Device (CCD) which greatly enhances the discrimination between fluorescence from the analyte and the background Raman scattering from the solvent. Register shifts between rows in the CCD are synchronized with the sample flow velocity so that fluorescence from a single species is collected in a single moving charge packet occupying an area approaching that of a single pixel while the background is spread evenly among a large number of pixels. This research has demonstrated that this technique is highly effective for the detection of fluorescently labelled latex microspheres. With additional development, the authors believe that this technique will achieve single molecule detection.