Patent ID: 11926822
Assignee: 10X GENOMICS, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method for determining a location of an analyte in three-dimensional space in a tissue sample, the method comprising:
(a) placing the tissue sample on an array and immobilizing the tissue sample by adding a hydrogel matrix to the tissue sample on the array, wherein the hydrogel matrix comprises a polymer;
(b) adding a plurality of spatially-programmed capture probes to the hydrogel matrix, wherein a spatially-programmed capture probe in the plurality of spatially-programmed capture probes comprises:
(i) a programmable migration domain that migrates to a z-axis location in the tissue sample;
(ii) a detectable moiety; and
(iii) a spatially-programmed capture domain that binds to the analyte;

(c) migrating the plurality of spatially-programmed capture probes into the hydrogel matrix from a point distal to a surface of the hydrogel matrix;
(d) hybridizing the spatially-programmed capture probe of the plurality of the spatially programmed capture probes to the analyte, thereby generating a hybridized spatially-programmed capture probe;
(e) detecting a z-axis location of the detectable moiety of the spatially programmed capture probe in the tissue sample, thereby determining the z-axis location of the spatially-programmed capture probe and analyte in the tissue sample;
(f) migrating the hybridized spatially-programmed capture probe to the array, wherein the array comprises a plurality of affixed capture probes, wherein an affixed capture probe of the plurality of affixed capture probes comprises a spatial barcode and a capture domain that hybridizes to the hybridized spatially-programmed capture probe; and
(g) determining (i) a sequence of the hybridized spatially-programmed capture probe, or a complement thereof, (ii) a sequence of the spatial barcode, or a complement thereof, and (iii) all or part of the sequence of the analyte, or a complement thereof, and using the determined sequences of (i), (ii), and (iii), and the determined z-axis location in (e), to identify the location of the analyte in the three-dimensional space in the tissue sample.