Pulsed-multiline excitation for color-blind fluorescence detection
First Claim
1. A pulse-multiline excitation apparatus for analyzing a sample containing one or more fluorescent species, comprising:
- one or more lasers configured to emit two or more excitation lines, each excitation line having a different wavelength;
a timing circuit coupled to the one or more lasers and configured to generate the two or more excitation lines sequentially according to a timing program that pulses the one or more lasers on and off in a firing sequence to produce time-correlated fluorescence emission signals from the sample;
a non-dispersive detector positioned to collect the time-correlated fluorescence emission signals emanating from the sample; and
an analyzer coupled to the detector and configured to associate the time-correlated fluorescence emission signals with the timing program to identify constituents of the sample.
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Abstract
The present invention provides a technology called Pulse-Multiline Excitation or PME. This technology provides a novel approach to fluorescence detection with application for high-throughput identification of informative SNPs, which could lead to more accurate diagnosis of inherited disease, better prognosis of risk susceptibilities, or identification of sporadic mutations. The PME technology has two main advantages that significantly increase fluorescence sensitivity: (1) optimal excitation of all fluorophores in the genomic assay and (2) “color-blind” detection, which collects considerably more light than standard wavelength resolved detection. Successful implementation of the PME technology will have broad application for routine usage in clinical diagnostics, forensics, and general sequencing methodologies and will have the capability, flexibility, and portability of targeted sequence variation assays for a large majority of the population.
100 Citations
38 Claims
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1. A pulse-multiline excitation apparatus for analyzing a sample containing one or more fluorescent species, comprising:
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one or more lasers configured to emit two or more excitation lines, each excitation line having a different wavelength; a timing circuit coupled to the one or more lasers and configured to generate the two or more excitation lines sequentially according to a timing program that pulses the one or more lasers on and off in a firing sequence to produce time-correlated fluorescence emission signals from the sample; a non-dispersive detector positioned to collect the time-correlated fluorescence emission signals emanating from the sample; and an analyzer coupled to the detector and configured to associate the time-correlated fluorescence emission signals with the timing program to identify constituents of the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38)
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Specification