Very large scale immobilized polymer synthesis
First Claim
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1. A method of detecting hybridization between biological polymers, comprising the acts of:
- providing a substrate having a surface including at least one biological polymer and at least one fluorescent label associated with the biological polymer;
generating an excitation laser beam;
scanning said laser beam relative to said surface;
collecting fluorescent radiation responsive to said laser beam using optics;
detecting said collected fluorescent radiation; and
autofocusing to bring into focus with respect to said optics at least a portion of said surface including the biological polymer.
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Abstract
A synthetic strategy for the creation of large scale chemical diversity. Solid-phase chemistry, photolabile protecting groups, and photolithography are used to achieve light-directed spatially-addressable parallel chemical synthesis. Binary masking techniques are utilized in one embodiment. A reactor system, photoremovable protecting groups, and improved data collection and handling techniques are also disclosed. A technique for screening linker molecules is also provided.
121 Citations
86 Claims
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1. A method of detecting hybridization between biological polymers, comprising the acts of:
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providing a substrate having a surface including at least one biological polymer and at least one fluorescent label associated with the biological polymer;
generating an excitation laser beam;
scanning said laser beam relative to said surface;
collecting fluorescent radiation responsive to said laser beam using optics;
detecting said collected fluorescent radiation; and
autofocusing to bring into focus with respect to said optics at least a portion of said surface including the biological polymer.
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2-62. -62. (canceled)
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63. A method of interrogating an addressable array unit having a transparent substrate with a back surface, and an array with a plurality of different chemical features on a front surface, the method comprising:
- (a) illuminating the features while the array is dry, with an interrogating light which is directed through the substrate from the back surface and onto the chemical features on the front surface; and
(b) detecting light emitted from respective features in response to the interrogating light, which detected light has passed from the front surface, through the substrate and out the back surface;
wherein the light is emitted from locations of the features which are spaced from the front surface a distance of less than one-eighth of the wavelength of the illuminating light in a gas or a vacuum which is in contact with the dry array. - View Dependent Claims (64, 65, 66, 67, 68, 69, 70, 71, 81)
- (a) illuminating the features while the array is dry, with an interrogating light which is directed through the substrate from the back surface and onto the chemical features on the front surface; and
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72. A method of interrogating an addressable array unit having a transparent substrate with a back surface, and an array with a plurality of different chemical features on a front surface, the method comprising:
- (a) illuminating the features while the array is dry, with an interrogating light which is directed through the substrate from the back surface and onto the chemical features on the front surface; and
(b) detecting light emitted from respective features in response to the interrogating light, which detected light has passed from the front surface, through the substrate and out the back surface;
wherein the light is emitted from locations of the features which are spaced from the front surface a distance of less than one-eighth of the wavelength of the emitted light in a gas or a vacuum which is in contact with the dry array. - View Dependent Claims (73, 74)
- (a) illuminating the features while the array is dry, with an interrogating light which is directed through the substrate from the back surface and onto the chemical features on the front surface; and
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75. A method of interrogating an addressable array unit having a substrate with a back surface, and an array with a plurality of different chemical features on a front surface, the method comprising:
- (a) illuminating the features while the array is dry, with an interrogating light which is directed through the substrate from the back surface and onto the chemical features on the front surface; and
(b) detecting light emitted from respective features in response to the interrogating light, which detected light has passed from the front surface, through the substrate and out the back surface;
wherein the light is emitted from locations of the features which are spaced from the front surface a distance such that the average detected signal from the dry array is at least 10% greater than would be detected under the same conditions except with the interrogating light and detected emitted light not passing through the substrate. - View Dependent Claims (76, 77)
- (a) illuminating the features while the array is dry, with an interrogating light which is directed through the substrate from the back surface and onto the chemical features on the front surface; and
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78. A package comprising:
- an addressable array unit having a transparent substrate with a back surface, and an array with a plurality of different chemical features on a front surface, the chemical features having a thickness of less than 100 nm; and
instructions to;
(i) interrogate the array with an interrogating light which is directed through the substrate from the back surface and onto the chemical features on the front surface; and
(ii) detect light emitted from respective features in response to the interrogating light, which detected light has passed from the front surface, through the substrate and out the back surface. - View Dependent Claims (79, 80)
- an addressable array unit having a transparent substrate with a back surface, and an array with a plurality of different chemical features on a front surface, the chemical features having a thickness of less than 100 nm; and
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82. A method for use with an interrogating an addressable array unit having a transparent substrate with a back surface, and an array with a plurality of different chemical features on a front surface, the method comprising:
- (a) machine reading an identifier associated with the array unit;
(b) based on the read identifier, retrieving by a processor an instruction that the array should be interrogated and read through the substrate from the back surface. - View Dependent Claims (83, 84, 85, 86)
- (a) machine reading an identifier associated with the array unit;
Specification