Randomly Ordered Arrays and Methods of Making and Using
Arrays including microparticles having probe and marker moieties are used for the detection of a target in a sample. Microparticles are randomly immobilized on at least a portion of a substrate. A detection scheme is performed to detect the marker associated with the microparticle and the identity of the probe, and any target bound to the probe.
1 . An array comprising:
a) a substrate;
b) a plurality of microparticles randomly immobilized on the wherein each microparticle comprises a self-encoding marker and a probe couple to the microparticle, wherein each self-encoding marker and probe comprises a unique self-encoding marker/probe pair, and wherein the probe is configured and arranged to specifically bind a target present in a sample; and
c) an immobilization material which immobilizes the microparticles on the substrate;
wherein the immobilization material comprises a reactive polymer, the reactive polymer bound to the substrate, and the microparticles immobilized on the substrate via the reactive polymer;
wherein the reactive polymer comprises a photoreactive polymer having at least one photoreactive group.
2 . The array of claim 1 wherein the photoreactive group is selected consisting of aryl ketones, arylazides, acyl azides, sulfonyl azides, phosphoryl azides, diazoalkanes, diazoketones, diazoacetates, and ketenes.
3 . The array of claim 1 the reactive polymer is selected from the group consisting of functionalized polyacrylamide, polymethacrylamide, polyvinylpyrrolidone, polyacrylic acid, polyethylene glycol, polyvinyl alcohol, poly(HEMA), copolymers thereof, and combinations thereof.
4 . The array of claim 1 wherein the reactive polymer is selected from the group consisting of photoreactive copolymers consisting of vinylpyrolidone and N-[3-(4-Benzoylbenzamido)propyl]methacrylamide (BBA-APMA); and acrylamide and BBA-APMA.
5 . The array of claim 1 wherein the reactive polymer comprises polymers selected from the group consisting of functionalized polysaccharides, glycosaminoglycans, polypeptides, and combinations thereof.
6 . The array of claim 1 wherein the immobilization material is patterned on the substrate thereby forming patches of immobilization material on the substrate.
7 . The array of claim 6 wherein the microparticles are immobilized on the substrate via the patches of immobilization material.
8 . The array of claim 7 wherein the plurality of microparticles comprises more than one subset of microparticles having unique self-encoding marker/probe pairs and the subsets of microparticles are individually disposed on separate patches of immobilization material on the substrate.
9 . The array of claim 1 wherein the microparticle further comprises a coating of a reactive compound, wherein the reactive compound is used to couple the microparticles to the immobilization material, the self-encoding marker, the probe, or any combination of the above.
10 . The array of claim 9 wherein the reactive compound includes reactive groups selected from the group consisting of carboxylic acids, sulfonic acids, phosphoric acids, phosphonic acids, aldehydes groups, amine groups, thiol groups, thiol-reactive groups, and epoxide groups.
11 . The array of claim 1 wherein the probe is a nucleic acid probe.
12 . The array of claim 1 wherein the probe is an antibody probe.
13 . The array of claim 1 wherein the self-encoding probe comprises at least one detectable particle and wherein the at least one detectable particle is coupled to the microparticle.
14 . The array of claim 13 wherein the detectable particle is selected from the group consisting of fluorphores, quantum dots, radioisotopes, and magnetic particles.
15 . The array of claim 14 wherein the self-encoding marker comprises a combination of detectable particles.