Method and apparatus for imaging a sample on a device
First Claim
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1. An apparatus for imaging a sample located on a support, said apparatus comprising:
- a body for immobilizing said support, said support comprising at least a first surface having said sample thereon;
an electromagnetic radiation source for generating excitation radiation having a first wavelength;
excitation optics for transforming the geometry of said excitation radiation to a line and directing said line at said sample for exciting a plurality of regions thereon, said line causing a labeled material on said sample to emit response radiation, said response radiation having a second wavelength, said first wavelength different from said second wavelength;
collection optics for collecting said response radiation from said plurality of regions;
a detector for sensing said response radiation received by said collection optics, said detector generating a signal proportional to the amount of radiation sensed thereon, said signal representing an image associated with said plurality of regions from said sample;
a translator coupled to said body for allowing a subsequent plurality of regions on said sample to be excited;
a processor for processing and storing said signal so as to generate a 2-dimensional image of said sample; and
a focuser for automatically focusing said sample in a focal plane of said excitation radiation.
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Abstract
Labeled targets on a support synthesized with polymer sequences at known locations according to the methods disclosed in U.S. Pat. No. 5,143,854 and PCT WO 92/10092 or others, can be detected by exposing selected regions of sample 1500 to radiation from a source 1100 and detecting the emission therefrom, and repeating the steps of exposition and detection until the sample is completely examined.
171 Citations
83 Claims
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1. An apparatus for imaging a sample located on a support, said apparatus comprising:
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a body for immobilizing said support, said support comprising at least a first surface having said sample thereon;
an electromagnetic radiation source for generating excitation radiation having a first wavelength;
excitation optics for transforming the geometry of said excitation radiation to a line and directing said line at said sample for exciting a plurality of regions thereon, said line causing a labeled material on said sample to emit response radiation, said response radiation having a second wavelength, said first wavelength different from said second wavelength;
collection optics for collecting said response radiation from said plurality of regions;
a detector for sensing said response radiation received by said collection optics, said detector generating a signal proportional to the amount of radiation sensed thereon, said signal representing an image associated with said plurality of regions from said sample;
a translator coupled to said body for allowing a subsequent plurality of regions on said sample to be excited;
a processor for processing and storing said signal so as to generate a 2-dimensional image of said sample; and
a focuser for automatically focusing said sample in a focal plane of said excitation radiation. - View Dependent Claims (2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 41)
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3. The apparatus as recited in claim 3 wherein said temperature controller comprises a thermoelectric cooler.
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25. A method for imaging a sample located on a support, said method comprising the steps of:
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immobilizing said support on a body;
exciting said sample on said support with an excitation radiation having a first wavelength from an electromagnetic radiation source, said excitation radiation having a linear geometry for exciting a plurality of regions on said sample;
detecting a response radiation having a second wavelength in response to said excitation radiation, said response radiation representing an image of said plurality of regions;
exciting a subsequent plurality of regions on said sample;
processing and storing said response radiation to generate a 2-dimensional image of said sample; and
auto-focusing said sample in a focal plane of said excitation radiation. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36)
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37. In a imaging system for imaging a sample on a support having a first surface and a second surface, said second surface being weakly reflective relative to said first surface, a method for focusing on said weakly reflective surface comprising the steps of:
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a) focusing said first surface;
b) focusing said second surface; and
c) finely focusing said second surface. - View Dependent Claims (38, 39, 40)
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42. A method, comprising the acts of:
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providing a substrate having a surface including biological polymers and at least one fluorescent label associated with at least one of the biological polymers;
providing a stage constructed to receive said substrate and being coupled to an autofocusing module;
generating an excitation laser beam;
scanning said laser beam relative to said surface;
collecting fluorescent radiation using optics, said fluorescent radiation being emitted from said at least one fluorescent label in response to said excitation beam;
detecting said collected fluorescent radiation; and
tilting said stage to focus said surface including the biological polymer with respect to said optics. - View Dependent Claims (43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57)
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58. A method of detecting hybridization between biological polymers, comprising the acts of:
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providing a flow cell including a light transparent substrate, said substrate having a surface located inside said flow cell, said surface including biological polymers and at least one fluorescent label associated with at least one of the biological polymers;
generating an excitation laser beam;
scanning said laser beam relative to said surface and delivering said laser beam to said biological polymer and said fluorescent label;
collecting fluorescent radiation emitted from said fluorescent label located inside said flow cell using optics; and
detecting said collected fluorescent radiation. - View Dependent Claims (59, 60, 61, 62, 63, 64, 65, 66, 78)
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67. A system for detecting hybridization between biological polymers, comprising:
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a laser source constructed to generate an excitation beam having a first wavelength;
a stage constructed to support a hybridization cell comprising a first surface including at least one biological polymer and at least one fluorescent label;
optics constructed to deliver said excitation beam to a region of said first surface to cause emission of fluorescent radiation from said fluorescent label;
a detector constructed and arranged to detect said fluorescent radiation and to generate a signal representing said detected fluorescent radiation;
a processor constructed and arranged to process said signal and generate an image corresponding to said fluorescent radiation detected over said first surface; and
an autofocuser, coupled to said stage, constructed and arranged to position said region of said first surface in focus with respect to said optics. - View Dependent Claims (68, 69, 70, 71, 72, 73, 74, 75, 76, 77)
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79. A system for detecting a hybridized sample, comprising:
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a laser source constructed and arranged to generate an excitation beam;
a stage constructed and arranged to support a cell having a first surface including a biological polymer and an associated fluorescent label located inside said cell;
optics constructed and arranged to deliver said excitation beam to a region of said first surface and cause emission of fluorescent radiation emitted from said label having a second wavelength different from said first wavelength;
a detector constructed and arranged to detect said fluorescent radiation emitted from said cell and generate a signal representing said fluorescent radiation; and
a processor constructed and arranged to process said signal and generate an image corresponding to said fluorescent radiation detected from said first surface. - View Dependent Claims (80, 81, 82, 83)
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Specification