| CPC G01N 33/505 (2013.01) [G01N 33/5052 (2013.01); G01N 33/5308 (2013.01); G01N 2333/70539 (2013.01)] | 11 Claims |
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1. A method comprising:
providing a substrate comprising a plurality of capture regions, wherein a capture region of the plurality of capture regions comprises:
a plurality of cell binding ligands affixed to the substrate, wherein a cell binding ligand of the plurality of cell binding ligands binds to a cell surface receptor, wherein each of the cell binding ligands is at least 100 microns from each other on the substrate; and
a plurality of nucleic acid barcode molecules affixed to the substrate, wherein each nucleic acid barcode molecule of the plurality of nucleic acid barcode molecules includes a common barcode sequence, wherein the common barcode sequence is unique to the capture region and an analyte binding sequence, wherein the plurality of the nucleic acid barcode molecules are within 1 to 100 microns of each cell binding ligand on the substrate;
contacting a plurality of cells each comprising the cell surface receptor with the substrate, wherein a subpopulation of the cells of the plurality of cells binds to the cell binding ligand of the plurality of cell binding ligands, thereby generating a captured cell subpopulation that binds the cell binding ligand at the capture region;
releasing a plurality of nucleic acid analytes from each of the captured cells of the captured subpopulation at the capture region to allow contact with the nucleic acid barcode molecules in proximity of the cell binding ligand and hybridizing the nucleic acid analytes of the plurality of nucleic acid analytes to the nucleic acid barcode molecules; and
forming a plurality of nucleic acid barcoded molecules from the plurality of nucleic acid analytes and the plurality of nucleic acid barcode molecules comprising extending the nucleic acid barcode molecule with a reverse transcriptase or a polymerase following hybridization,
wherein each formed nucleic acid barcoded molecule of the plurality of nucleic acid barcoded molecules includes the common barcode sequence, or a complement thereof, and a sequence corresponding to the nucleic acid analyte, or a complement thereof.
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