METHOD OF MAKING AND USING MICROARRAYS SUITABLE FOR HIGH THROUGHPUT DETECTION
First Claim
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1. A method of making a microarray, comprising:
- a. providing a substrate that is at least translucent having at least an upper surface and a lower surface;
b. depositing a cladding layer material onto the upper surface of the substrate so as to create a cladding layer on the substrate;
c. etching an array of sensing zones into the cladding layer, each sensing zone being a discrete area in which the cladding layer has been removed to expose the underlying substrate;
d. applying to the array of sensing zones a ligand having an apical functional group and a photolabile functional group, wherein the photolabile functional group requires radiant light energy in order to bind to a surface; and
e. irradiating the lower surface of the substrate with radiant light energy so that the radiant light energy passes through the upper surface only in the sensing zones, thereby causing the ligand in the sensing zones to bind to the upper surface;
wherein the bound ligand is substantially confined to the sensing zones and the cladding layer remains substantially free of bound ligand.
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Abstract
Disclosed are high density microarrays and methods for making and using such microarrays. The microarrays of the present invention can have uniformly shaped and sized sensing zones and are designed to allow high-throughput detection assays with minimal noise.
38 Citations
42 Claims
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1. A method of making a microarray, comprising:
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a. providing a substrate that is at least translucent having at least an upper surface and a lower surface; b. depositing a cladding layer material onto the upper surface of the substrate so as to create a cladding layer on the substrate; c. etching an array of sensing zones into the cladding layer, each sensing zone being a discrete area in which the cladding layer has been removed to expose the underlying substrate; d. applying to the array of sensing zones a ligand having an apical functional group and a photolabile functional group, wherein the photolabile functional group requires radiant light energy in order to bind to a surface; and e. irradiating the lower surface of the substrate with radiant light energy so that the radiant light energy passes through the upper surface only in the sensing zones, thereby causing the ligand in the sensing zones to bind to the upper surface;
wherein the bound ligand is substantially confined to the sensing zones and the cladding layer remains substantially free of bound ligand. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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16. A method of making a microarray, comprising:
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a. depositing a cladding layer material onto a substrate so as to create a cladding layer on the substrate; b. etching an array of sensing zones into the cladding layer, each sensing zone having a shape and being located at a discrete area in which the cladding layer has been removed to expose the underlying substrate; and c. applying a ligand having a basal functional group and an apical functional group to the array of sensing zones, wherein the basal functional group is configured to bind to the substrate but not to the cladding layer material, so that bound ligand is substantially confined to the sensing zones and the cladding layer remains substantially free of bound ligand. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
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27. A method of making a microarray, comprising:
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a. depositing a cladding layer material onto a substrate so as to create a cladding layer on the substrate; b. patterning an array of sensing zones onto the cladding layer; c. removing the cladding layer material outside of the sensing zones to expose the underlying substrate, so that each sensing zone comprises a discrete area of cladding layer material surrounded by exposed substrate; and d. applying a ligand having a basal functional group and an apical functional group to the array of sensing zones, wherein the basal functional group is configured to bind to the cladding layer material but not to the substrate material, so that bound ligand is substantially confined to the sensing zones and the substrate remains substantially free of bound ligand. - View Dependent Claims (28, 29, 30, 31, 32, 33, 34, 35, 36, 37)
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38. A microarray, comprising:
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a. a substrate; b. cladding material deposited on the substrate in the form of an array of discontinuous islands of cladding material substantially isolated from one another by the substrate; and c. a ligand having a basal functional group and an apical functional group attached to the islands, wherein the basal functional group is configured to bind to the cladding material but not to the substrate so that bound ligand is substantially confined to the islands and the substrate remains substantially free of bound ligand.
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39. A microarray, comprising:
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a. a substrate; b. cladding material deposited on the substrate in the form of a continuous layer of cladding material that laterally defines an array of discontinuous wells, wherein a base portion of said wells includes the substrate; and c. a ligand having a basal functional group and an apical functional group attached to the wells, wherein the basal functional group is configured to bind to the substrate but not to the cladding layer material so that bound ligand is substantially confined to the sensing zones and the cladding layer remains substantially free of bound ligand. - View Dependent Claims (40, 41, 42)
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Specification