Identifying randomly distributed microparticles in images to sequence a polynucleotide
First Claim
1. A computer-implemented method for performing sequencing of a polynucleotide, comprising:
- receiving, by the computer, a first image of microparticles, wherein the first image is stored in a memory of the computer, wherein the microparticles are distributed in a random fashion on a substrate wherein the microparticles have respective random positions on the substrate, wherein the respective random positions are not pre-determined and are not regularly spaced, and wherein each of the microparticles comprises a plurality of similar oligonucleotides of the polynucleotide;
receiving, by the computer, a second image of the microparticles, wherein the second image of the microparticles highlights at least some of the microparticles in response to fluorescent chemicals attached to the oligonucleotides based on terminal nucleotides of the oligonucleotides on the microparticles, and wherein the second image is stored in the memory of the computer;
determining, by the computer, a plurality of first subportions of the first image, wherein each subportion comprises a respective plurality of microparticles distributed in a random fashion, and wherein each first subportion'"'"'s random distribution comprises a respective unique pattern;
analyzing, by the computer, the second image to identify a plurality of second subportions in the second image that each corresponds to a respective one of the plurality of first subportions based on the respective unique patterns;
matching, by the computer, a plurality of the microparticles in the first and second images based on said analyzing using the respective unique patterns; and
determining, by the computer, the sequence of nucleotides of the polynucleotide based on said matching and the highlighted microparticles, wherein the sequence of nucleotides is stored in the memory of the computer.
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Abstract
Performing sequencing of a polynucleotide. A first image of microparticles that are distributed in a random fashion on a substrate may be received. Each of the microparticles may include a plurality of similar oligonucleotides of the polynucleotide. A second image of the microparticles may be received. A plurality of first subportions of the first image may be determined. Each subportion may include a respective plurality of microparticles distributed in a random fashion. The second image may be analyzed to identify a plurality of second subportions in the second image. Each of the plurality of second subportions may correspond to a respective one of the plurality of first subportions. A plurality of the microparticles may be matched from the first and second images based on said analyzing. At least a portion of the sequence of nucleotides of the polynucleotide may be determined based on said matching.
62 Citations
25 Claims
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1. A computer-implemented method for performing sequencing of a polynucleotide, comprising:
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receiving, by the computer, a first image of microparticles, wherein the first image is stored in a memory of the computer, wherein the microparticles are distributed in a random fashion on a substrate wherein the microparticles have respective random positions on the substrate, wherein the respective random positions are not pre-determined and are not regularly spaced, and wherein each of the microparticles comprises a plurality of similar oligonucleotides of the polynucleotide; receiving, by the computer, a second image of the microparticles, wherein the second image of the microparticles highlights at least some of the microparticles in response to fluorescent chemicals attached to the oligonucleotides based on terminal nucleotides of the oligonucleotides on the microparticles, and wherein the second image is stored in the memory of the computer; determining, by the computer, a plurality of first subportions of the first image, wherein each subportion comprises a respective plurality of microparticles distributed in a random fashion, and wherein each first subportion'"'"'s random distribution comprises a respective unique pattern; analyzing, by the computer, the second image to identify a plurality of second subportions in the second image that each corresponds to a respective one of the plurality of first subportions based on the respective unique patterns; matching, by the computer, a plurality of the microparticles in the first and second images based on said analyzing using the respective unique patterns; and determining, by the computer, the sequence of nucleotides of the polynucleotide based on said matching and the highlighted microparticles, wherein the sequence of nucleotides is stored in the memory of the computer. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A non-transitory, computer accessible memory medium storing program instructions for performing sequencing of a polynucleotide, wherein the program instructions are executable to:
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receive a first image of microparticles, wherein the microparticles are distributed in a random fashion on a substrate wherein the microparticles have respective random positions on the substrate, wherein the respective random positions are not pre-determined and are not regularly spaced, and wherein each of the microparticles comprise a plurality of similar oligonucleotides of the polynucleotide; receive a second image of the microparticles, wherein the second image of the microparticles highlights at least some of the microparticles in response to fluorescent chemicals attached to the oligonucleotides based on terminal nucleotides of the oligonucleotides on the microparticles; determine a plurality of first subportions of the first image, wherein each subportion comprises a respective plurality of microparticles distributed in a random fashion, and wherein each first subportion'"'"'s random distribution comprises a respective unique pattern; analyze the second image to identify a plurality of second subportions in the second image that each corresponds to a respective one of the plurality of first subportions based on the respective unique patterns; match a plurality of the microparticles in the first and second images based on said analyzing using the respective unique patterns; and determine the sequence of nucleotides of the polynucleotide based on said matching and the highlighted microparticles. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20)
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21. A system for performing sequencing of a polynucleotide, comprising:
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an image acquisition apparatus, wherein the image acquisition apparatus is configured to acquire images of microparticles, wherein the microparticles are distributed in a random fashion on a substrate wherein the microparticles have respective random positions on the substrate, wherein the respective random positions are not pre-determined and are not regularly spaced, and wherein each of the microparticles comprise a plurality of similar oligonucleotides of the polynucleotide; a processor coupled to the image acquisition device; a memory medium coupled to the processor, wherein the memory medium stores program instructions that are executable by the processor to; receive a first image of microparticles via the image acquisition apparatus at a first time; receive a second image of the microparticles via the image acquisition apparatus at a second time wherein the second image of the microparticles highlights at least some of the microparticles in response to fluorescent chemicals attached to the oligonucleotides based on terminal nucleotides of the oligonucleotides on the microparticles; determine a plurality of first subportions of the first image, wherein each subportion comprises a respective plurality of microparticles distributed in a random fashion, and wherein each first subportion'"'"'s random distribution comprises a respective unique pattern; analyze the second image to identify a plurality of second subportions in the second image that each corresponds to a respective one of the plurality of first subportions based on the respective unique patterns; match a plurality of the microparticles in the first and second images based on said analyzing using the respective unique patterns; and determine the sequence of nucleotides of the polynucleotide based on said matching and the highlighted microparticles. - View Dependent Claims (22, 23, 24, 25)
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Specification