Multiplexed capillary electrophoresis system
First Claim
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1. A method for detecting fluorescent target species in a sample using multiplexed capillary electrophoresis, comprising:
- (a) providing a capillary array comprising a plurality of coplanar parallel capillaries, each capillary having an intake end, an outflow end, and an annular wall with a first transparent portion defining a transparent path extending through the capillary array perpendicular to the capillaries;
(b) introducing a sample containing a fluorescent target species into the intake end of at least one capillary such that the sample migrates through the capillary toward the outflow end;
(c) inducing fluorescence emission from the target species by irradiating the species with a beam of coherent light having a wavelength of about 200-1500 nm directed along the transparent path; and
(d) detecting fluorescence emission from the target species.
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Abstract
The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.
267 Citations
59 Claims
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1. A method for detecting fluorescent target species in a sample using multiplexed capillary electrophoresis, comprising:
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(a) providing a capillary array comprising a plurality of coplanar parallel capillaries, each capillary having an intake end, an outflow end, and an annular wall with a first transparent portion defining a transparent path extending through the capillary array perpendicular to the capillaries; (b) introducing a sample containing a fluorescent target species into the intake end of at least one capillary such that the sample migrates through the capillary toward the outflow end; (c) inducing fluorescence emission from the target species by irradiating the species with a beam of coherent light having a wavelength of about 200-1500 nm directed along the transparent path; and (d) detecting fluorescence emission from the target species. - View Dependent Claims (2, 3, 4, 5, 6)
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7. A method for detecting fluorescent target species in a sample using capillary electrophoresis comprising:
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(a) providing a capillary array having a plurality of coplanar parallel capillaries, each capillary having an intake end, an outflow end, and an annular wall, each annular wall defining an interior portion and having a transparent portion for optically coupling the interior portion to an image array detector; (b) optically coupling the capillary array to an image array detector having linearly aligned pixels located in a plane parallel to the capillary array such that at least one of the capillaries is optically coupled to a plurality of pixels; (c) introducing a sample containing a fluorescent target species into the intake end of the at least one of the capillaries such that the sample migrates through the capillary toward the outflow end; (d) inducing fluorescence emission from the target species by irradiating the species with a beam of coherent light; and (e) detecting the fluorescence emission through the transparent portion of the capillary using the optically coupled pixels. - View Dependent Claims (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)
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36. A method for detecting a fluorescent DNA fragment in a sample using multiplexed capillary electrophoresis, comprising:
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(a) providing a capillary array of coplanar parallel capillaries, each capillary having an intake end, an outflow end, and an annular wall defining an interior portion, each annular wall having a transparent portion optically coupled to a detector; (b) introducing a sample containing a DNA fragment into the intake end of at least one capillary such that the sample migrates through the capillary toward the outflow end; (c) inducing fluorescence emission from the DNA fragment by irradiating the sample with a single beam of coherent light; (d) splitting the fluorescence emission into a first emission channel and a second emission channel using a first long-pass filter and a second long-pass filter, respectively; and (e) detecting fluorescence emission in the first and second emission channels, wherein detection is effected through the transparent portion of the annular wall. - View Dependent Claims (37, 38, 39, 40, 41, 42, 43, 44, 45, 46)
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47. A method for detecting fluorescent target species in a sample using capillary electrophoresis comprising:
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(a) providing at least one capillary having an intake end, an outflow end, and an annular wall, each annular wall defining an interior portion and having a transparent portion for optically coupling the interior portion to a detector; (b) optically coupling the at least one capillary to the detector; (c) introducing a sample containing a fluorescent target species into the intake end of the at least one capillary such that the sample migrates through the capillary toward the outflow end; (d) inducing fluorescence emission from the target species by irradiating the species with a beam of coherent light having a wavelength of about 200-1500 nm; (e) splitting the fluorescence emission into first and second emission channels; and (f) simultaneously detecting the fluorescence emission in the first and second emission channels through the transparent portion of the capillary. - View Dependent Claims (48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59)
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Specification