CPC C12Q 1/6818 (2013.01) [C12N 5/0062 (2013.01); C12Q 1/6804 (2013.01); C12Q 1/6837 (2013.01); C12Q 1/6841 (2013.01); C12Q 1/6876 (2013.01); G01N 33/5023 (2013.01); G01N 33/5088 (2013.01); G01N 33/54306 (2013.01); C12Q 2600/158 (2013.01); G01N 2458/10 (2013.01)] | 30 Claims |
1. A method for identifying location and/or abundance of an analyte in a plurality of live cells, the method comprising:
(a) contacting the plurality of live cells with a substrate comprising a plurality of capture probes, wherein a capture probe of the plurality of capture probes comprises (i) a spatial barcode and (ii) a capture domain that binds to a sequence present in the analyte;
(b) perfusing one or more test compounds through a perfusion chamber comprising a gasket and the substrate, wherein the gasket comprises (i) a plurality of apertures, (ii) a plurality of input channels connected to the plurality of apertures, and (iii) a plurality of output channels connected to the plurality of apertures, and wherein the plurality of live cells is located within the perfusion chamber;
(c) hybridizing the analyte to the capture domain of the capture probe;
(d) extending the capture probe using the analyte as a template, thereby generating an extended capture probe;
(e) amplifying the extended capture probe to produce a plurality of extended capture probes; and
(f) sequencing (i) the spatial barcode, or a complement thereof, and (ii) all or part of the sequence of the analyte, or a complement thereof, and using the determined sequences of (i) and (ii) to identify the location and/or abundance of the analyte in the plurality of live cells.
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