Method in inspecting DNA and apparatus therefor
First Claim
Patent Images
1. A method of obtaining fluorescent images of samples of fluorescent material, comprising the steps of:
- (a) moving a substrate in a first direction, said substrate comprising a an array of compartments for carrying said samples, each row in the array of compartments being seperated by an equal pitch distance;
(b) sequentially irradiating said compartments in said array of compartments with an exciting light while moving said substrate in said first direction by scanning said exciting light over said pitch distance at substantially the same speed as the moving substrate, including varying an intensity of said exciting light from a first irradiating value to a second irradiating value during the period of time that one compartment is scanned;
(c) detecting fluorescent light generated from each of said samples due to irradiation by said exciting light while cutting off light reflected from said substrate due to the scanning of said exciting light; and
(d) determining a value of fluorescence as generated from each of said samples based upon information from said fluorescent light detected in step (c).
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Abstract
A method and apparatus of stably obtaining fluorescent images of two or more fluorescent samples includes disposing the samples onto compartments defined on a substrate. The compartments are sequentially irradiated with an exciting light where the intensity varies. A value of fluorescence as generated from each of the samples is determined, and a fluorescent image is obtained based on the value of fluorescence.
21 Citations
18 Claims
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1. A method of obtaining fluorescent images of samples of fluorescent material, comprising the steps of:
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(a) moving a substrate in a first direction, said substrate comprising a an array of compartments for carrying said samples, each row in the array of compartments being seperated by an equal pitch distance; (b) sequentially irradiating said compartments in said array of compartments with an exciting light while moving said substrate in said first direction by scanning said exciting light over said pitch distance at substantially the same speed as the moving substrate, including varying an intensity of said exciting light from a first irradiating value to a second irradiating value during the period of time that one compartment is scanned; (c) detecting fluorescent light generated from each of said samples due to irradiation by said exciting light while cutting off light reflected from said substrate due to the scanning of said exciting light; and (d) determining a value of fluorescence as generated from each of said samples based upon information from said fluorescent light detected in step (c). - View Dependent Claims (2, 3)
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4. A method of obtaining fluorescent images of samples which may include a fluorescent material, comprising steps of:
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(A) moving a substrate in a first direction, said substrate comprising an array of compartments, each row in the array of compartments being separated by an equal pitch distance and each compartment disposed with a sample; (B) sequentially irradiating said samples with an exciting light while moving said substrate by scanning the exciting light in said first direction over said pitch distance at substantially the same speed as th moving substrate, wherein an intensity of said exciting light is changed from a first irradiating value to a second irradiating value within a period of time during which each sample is irradiated; (C) obtaining fluorescent images of each of said samples by detecting fluorescent lights generated from each of said samples due to the sequential irradiation of said samples by the exciting light performed in said irradiating step (B) while cutting off exciting light reflected from said substrate; and (D) processing said fluorescent images of each of said samples as obtained in said obtaining step (C) so as to determine a true value of fluorescence as generated from each of said samples. - View Dependent Claims (5, 6, 7)
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8. A method of inspecting DNA, comprising the steps of:
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(a) moving a substrate in a first direction, the substrate containing a plurality of DNA-samples arranged in an array of rows separated by an equal pitch distance, wherein each DNA-sample is labeled with a fluorescence and each DNA-sample is disposed on a bead; (b) sequentially irradiating each of p DNA-samples in sad plurality of DNA-samples with q exciting lights by scanning said exciting lights over said pitch distance at substantially the same steed as the moving substrate, wherein p≧
2;(c) switching each of said exciting lights into r beams different in intensity from one another within a scanning period during which each of said DNA-samples is irradiated, wherein r≧
2;(d) detecting r fluorescent lights as generated from each of said DNA-samples in said irradiating step (a), corresponding to said beams while cutting off light reflected from said substrate due to irradiation of said substrate by said q exciting lights; (e) inspecting a DNA in each of said DNA-samples by using information upon said r fluorescent lights each different in intensity from one another as detected in said detecting step (b). - View Dependent Claims (9, 10)
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11. A method of inspecting DNA, comprising the steps of:
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(a) moving a substrate in a first direction, the substrate containing a plurality of DNA-samples arrange an array of rows separated by an equal pitch distance, wherein each DNA-sample is labeled with a fluorescence and and each DNA-sample is disposed on a bead; (b) sequentially irradiating each of p DNA-samples in said plurality of DNA-samples with q exciting lights by scanning said q exciting lights over said pitch distance at substantially the same speed as the moving substrate, wherein p≧
2;(c) switching each of said q exciting lights into two beams different in intensity from each other within a scannig period during which each of said DNA-samples is irradiated; (d) converging two fluorescent lights as generated from each of said DNA-samples in said irradiating step (b), corresponding to said two beams different in intensity from each other while cutting off light reflected from said substrate due to irradiation of said substrate by said exciting lights; (e) branching said two fluorescent lights as converged in said converging step (d) into two fluorescent lights different in intensity from each other so as to detect each of said branched two fluorescent lights; and (f) inspecting a DNA in each of said DNA-samples by using information upon each of said two fluorescent lights as detected in said detecting step (e). - View Dependent Claims (12, 13, 14)
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15. A method of inspecting DNA, comprising the steps of:
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(a) continuously moving, in one direction, p DNA-samples labeled with a fluorescence, each DNA-sample disposed on the surface of an array of beads or dots arranged at a fixed pitch on a substrate, wherein p≧
2;(b) sequentially irradiating each of said beads or dots with one spot of a spot exciting-light comprising one or more spots, each of said one or more spots of said spot exciting-light having a beam diameter approximately equal to a diameter of said beads or dots, wherein said beads or dots are moved in one direction and are irradiated with said spots of said spot exciting-light and said spot exciting-light moves in said one direction at substantially the same speed as said beads or dots so that said spot exciting-light tracks said beads or dots, wherein each of said beads or dots is irradiated by a period of time substantially equal to the time required for moving said spots of said spot exciting-light by said fixed pitch, and wherein each spot of said spot exciting-light changes intensity from a first irradiating value to a second irradiating value within said period of time during which said beads are irradiated; (c) separating fluorescence as generated from each of said DNA-samples from said spot exciting-light so as to detect said fluorescence while cutting off light reflected from said substrate due to irradiation of said substrate by said spot of exciting-light; and (d) inspecting said DNA samples by using information as obtained in said detecting step (c). - View Dependent Claims (16, 17, 18)
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Specification