Methods and compositions related to microsphere surface gelation
Assignee10x Genomics, Inc.
InventorsShalini Gohil, Shea Thompson Lance, Geoffrey P. McDermott, David Michael Patterson, Andrew D. Price
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Abstract
The present disclosure relates generally to compositions and methods for generating hydrogel coatings on features. In some embodiments, hydrogel-coated features are prepared by horseradish peroxidase-mediated gelation of microspheres. Some embodiments relate to methods for generating hydrogel coatings using particle templated emulsification. The disclosure also relates to the use of hydrogel-coated features in arrays for spatial analysis of biological analytes in biological samples.
Claims22
A hydrogel-coated feature comprising:
- (a) a solid particle;
- and
- (b) a hydrogel encapsulating the solid particle, wherein the hydrogel comprises a crosslinked polymer and a plurality of moieties.
A method of preparing a hydrogel-coated feature, comprising:
- (a) providing a solid particle;
- and
- (b) encapsulating the solid particle with a hydrogel comprising a crosslinked polymer and a plurality of a moieties.
A method for preparing a plurality of hydrogel-coated features comprising:
- (a) suspending a plurality of features in an aqueous solution comprising a linear polymer, a surfactant, and an enzyme, thereby forming a suspension comprising the plurality of features;
- (b) adding an oil to the suspension and emulsifying the suspension into a plurality of droplets, wherein a droplet of the plurality of droplets comprises a feature of the plurality of features coated with the linear polymer and the enzyme;
- (c) contacting the plurality of droplets with a substrate for the enzyme, wherein the enzyme mediates crosslinking of the linear polymer to form a hydrogel on a feature of the plurality of features, thereby forming a plurality of hydrogel-coated features.
A method of immobilizing a capture probe on a feature, comprising:
- (a) encapsulating a solid particle with a hydrogel comprising a crosslinked polymer, and
- (b) attaching the capture probe to a moiety within the hydrogel.
A method for preparing a plurality of hydrogel-coated features comprising:
- (a) suspending a plurality of features in an aqueous solution comprising a polymerizable monomer and a surfactant, thereby forming a suspension comprising the plurality of features;
- (b) adding a partitioning oil to the suspension and emulsifying the suspension into a plurality of droplets, wherein a droplet of the plurality of droplets comprises a feature of the plurality of features coated with the monomer;
- (c) contacting the plurality of droplets with a polymerization agent such that the monomer coating a feature of the plurality of features is polymerized to form a plurality of hydrogel-coated features.
Dependencies are inferred from the claim text.