Methods and compositions for multiplex cell analysis
Assignee10x Genomics, Inc.
InventorMichael Schnall-Levin
Life of this patent
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Abstract
Provided herein in some aspects are methods, compositions, kits, and systems for performing multiplexed single-cell analysis on an in situ platform, providing alternatives which have a higher cell throughput and/or lower cost per cell compared to current single-cell analysis techniques. In some embodiments, the methods disclosed herein comprise using labeling agents that comprise sample-specific barcodes and/or cell feature specific barcodes (e.g., analyte specific barcodes) to label single-cell populations, immobilizing the labeled cells, and performing in situ detection of the labeling agents and/or other features including cellular analytes of the labeled cells.
Claims20
A method for sample analysis, comprising:
- (a) contacting a first sample of dissociated cells with a first labeling agent and a second sample of dissociated cells with a second labeling agent,
- wherein the first labeling agent and the second labeling agent each comprises (i) a binding moiety that binds to cells of the first sample and cells of the second sample, respectively, and (ii) a reporter oligonucleotide comprising a first barcode region comprising a sample-specific barcode sequence that identifies the sample origin of the cells as the first sample and the second sample, respectively
- (b) combining cells from the first sample that are labeled with the first labeling agent and cells from the second sample that are labeled with the second labeling agent to provide combined labeled cells;
- (c) immobilizing at least a portion of the combined labeled cells on a substrate to provide immobilized dissociated cells;
- (d) contacting the immobilized dissociated cells with
- a probe that directly or indirectly binds to an analyte in one or more of the immobilized dissociated cells, wherein the probe comprises a second barcode region that identifies the analyte;
- and
- (e) analyzing (i) the sample-specific barcode sequence or a complement thereof and (ii) the second barcode region or a complement thereof in the immobilized dissociated cells on the substrate using nucleic acid sequencing or probe hybridization.
Dependencies are inferred from the claim text.