Multiplexed fluorescence detector system for capillary electrophoresis
First Claim
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1. A multiplexed fluorescence detection system for use with capillary electrophoresis to detect target species in a sample, comprising:
- (a) an array of at least two capillaries each having an intake end, an outflow end, and an annular wall wherein at least a portion of the wall is translucent to light having a wavelength of about 200-1,500 nm, the intake end being in fluid communication with the sample so that the sample is drawn into the capillary;
(b) an array of at least two optical fibers, each optical fiber having a first and second end, the second end of each optical fiber being coupled to a capillary in the array of capillaries;
(c) means for generating coherent light positioned to direct the coherent light onto the first end of the optical fibers, wherein the coherent light is of a wavelength suitable to cause fluorescence in target species present in the capillaries; and
(d) means for substantially simultaneously detecting changes in fluorescent emission by the target species through the annular wall of each capillary.
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Abstract
A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis.
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Citations
37 Claims
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1. A multiplexed fluorescence detection system for use with capillary electrophoresis to detect target species in a sample, comprising:
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(a) an array of at least two capillaries each having an intake end, an outflow end, and an annular wall wherein at least a portion of the wall is translucent to light having a wavelength of about 200-1,500 nm, the intake end being in fluid communication with the sample so that the sample is drawn into the capillary; (b) an array of at least two optical fibers, each optical fiber having a first and second end, the second end of each optical fiber being coupled to a capillary in the array of capillaries; (c) means for generating coherent light positioned to direct the coherent light onto the first end of the optical fibers, wherein the coherent light is of a wavelength suitable to cause fluorescence in target species present in the capillaries; and (d) means for substantially simultaneously detecting changes in fluorescent emission by the target species through the annular wall of each capillary. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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17. A multiplexed method for detecting the presence of fluorescent target species in a sample in a plurality of capillaries using capillary electrophoresis, comprising the steps of:
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(a) introducing samples containing a fluorescent species into intake ends of a plurality of capillaries, each capillary having an annular wall, and an outflow end, wherein the samples migrate through the capillaries towards the outflow ends; (b) irradiating the samples in the capillaries with coherent light transmitted through optical fibers, the optical fibers having first ends coupled to a coherent light source and second ends individually coupled to the capillaries; (c) substantially simultaneously detecting change in fluorescent emission by the fluorescent species through the wall of each of the capillaries. - View Dependent Claims (18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28)
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29. A multiplexed method of detecting the presence of macromolecule in a sample present in a plurality of capillaries in capillary electrophoresis, comprising the steps of:
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(a) linking a fluorophore to the macromolecule in a sample to transform the macromolecule into a fluorescent species with fluorescence that can be excited by light of suitable wavelength; (b) introducing samples containing fluorescent species into each of a plurality of capillaries, each capillary having an annular wall, an intake end and an outflow end; (c) irradiating each sample in each capillary with coherent light to cause fluorescence of the macromolecule, the coherent light being transmitted though optical fibers individually coupled to the capillaries; and (d) substantially simultaneously detecting changes in fluorescent emission through the wall of each of the capillaries by the fluorescent species using a charge transfer device based imaging system. - View Dependent Claims (30, 31, 32, 33, 34, 35)
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36. A multiplexed method of detecting the presence of macromolecules in samples present in a plurality of capillaries in capillary electrophoresis, comprising the steps of:
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(a) providing a buffer solution containing a fluorophore; (b) introducing samples into a plurality of capillaries in a capillary electrophoresis system, each capillary containing the buffer; (c) irradiating the samples in the capillaries with coherent light to cause fluorescence, the coherent light being transmitted though optical fibers individually coupled to the capillaries; and (d) substantially simultaneously detecting changes in fluorescent emission through the wall of each the capillaries by the fluorescent species by using a charge transfer device based imaging system.
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37. A multiplexed method of detecting the presence of DNA fragments in samples present in a plurality of capillaries in capillary electrophoresis, comprising the steps of:
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(a) linking a fluorophore to a DNA fragment in a sample to transform it into a fluorescent species; (b) introducing samples containing fluorescent species into a plurality of capillaries in a capillary electrophoresis system, the system comprises an array of at least about 100 capillaries each having an inside diameter of about 20-500 microns, and an annular wall; (c) irradiating the samples in the capillaries with coherent light having a wavelength of about 250-700 nm to cause native fluorescence of the DNA fragment, the coherent light being transmitted though optical fibers individually coupled to the capillaries; and (d) substantially simultaneously detecting change in fluorescence emitted by the fluorescent species through the wall of each of the capillaries by focusing the fluorescence onto a charge transfer device based imaging system.
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Specification