High performance spatial mapping of individual targets using releasable handshake sequences

Abstract

Systems, methods, and compositions for generating a spatial map of a distribution of targets of a sample are described. A system for mapping targets can include a substrate; and a distribution of functionalized features associated with the substrate, wherein a representative feature of the distribution of functionalized features comprises: one or more functionalized molecules coupled to the representative feature, the one or more molecules including at least: a handshake sequence comprising a reactive portion, a barcode segment serving as a spatial address, and a cleavable linker configured to allow the handshake sequence to be released from the representative feature in response to a stimulus. The system can be used to tag nuclei, cells, and/or other target components of a sample.