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US 2021/0332425 A1Published application
Increasing spatial array resolution
Assignee10x Genomics, Inc.
InventorsKatherine Pfeiffer, Rajiv Bharadwaj, Stephane Boutet, Lucas Frenz
Life of this patent
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Abstract
This disclosure relates to methods of manufacturing of spatial arrays and determining the location of an analyte present in a biological sample (e.g., by utilizing a cell-tagging agent).
Claims56
A method for determining the location of an analyte present in a biological sample, comprising:
- (a) providing a substrate comprising an arrayed plurality of spatially-barcoded oligonucleotides, wherein a spatially-barcoded oligonucleotide of the plurality of spatially-barcoded oligonucleotides comprises a spatial barcode, a priming domain, and a first hybridization domain;
- (b) coupling a cell-tagging agent to the spatially-barcoded oligonucleotide;
- (c) contacting the biological sample to the cell-tagging agent such that a cell in the biological sample is tagged with the spatially barcoded-oligonucleotide;
- (d) providing the cell comprising the spatially barcoded-oligonucleotide to a bead,
- wherein the bead comprises:
- 1. a first bead-bound oligonucleotide, wherein the first bead-bound oligonucleotide comprises a cellular barcode and a second hybridization domain, and
- 2. a second bead-bound oligonucleotide, wherein the second bead-bound oligonucleotide comprises the cellular barcode and a capture domain;
- (e) allowing an analyte from the cell to interact with the capture domain of the second bead-bound oligonucleotide;
- (f) associating the analyte bound to the capture domain of the second bead-bound oligonucleotide with the cellular barcode;
- and
- (g) associating the cellular barcode with the spatial barcode, thereby determining the location of the analyte present in the biological sample.
8 more elements
A method for determining the location of an analyte present in a biological sample, comprising:
- (a) providing a substrate comprising an arrayed plurality of spatially-barcoded oligonucleotides, wherein a spatially-barcoded oligonucleotide of the plurality of spatially-barcoded oligonucleotides comprises a spatial barcode, a first hybridization domain, and a cleavage domain;
- (b) coupling a cell-tagging agent with the spatially-barcoded oligonucleotide;
- and
- (c) determining the location of an analyte present in the biological sample.
1 more element
A method for generating an array, comprising:
- (a) providing a plurality of spatially-barcoded oligonucleotides to a substrate, wherein two or more spatially-barcoded oligonucleotides of the plurality of spatially-barcoded oligonucleotides comprise a first attachment sequence and a second attachment sequence and wherein the substrate comprises a plurality of functional domains, wherein at least two functional domains of the plurality of functional domains hybridizes to the at least two spatially-barcoded oligonucleotides, thus generating an array of spatially-barcoded oligonucleotides;
- and
- (b) amplifying the spatially-barcoded oligonucleotides on the substrate, thereby generating the array.
Dependencies are inferred from the claim text.