PT - JOURNAL ARTICLE AU - Christopher R. Merritt AU - Giang T. Ong AU - Sarah Church AU - Kristi Barker AU - Gary Geiss AU - Margaret Hoang AU - Jaemyeong Jung AU - Yan Liang AU - Jill McKay-Fleisch AU - Karen Nguyen AU - Kristina Sorg AU - Isaac Sprague AU - Charles Warren AU - Sarah Warren AU - Zoey Zhou AU - Daniel R. Zollinger AU - Dwayne L. Dunaway AU - Gordon B. Mills AU - Joseph M. Beechem TI - High multiplex, digital spatial profiling of proteins and RNA in fixed tissue using genomic detection methods AID - 10.1101/559021 DP - 2019 Jan 01 TA - bioRxiv PG - 559021 4099 - http://biorxiv.org/content/early/2019/02/26/559021.short 4100 - http://biorxiv.org/content/early/2019/02/26/559021.full AB - We have developed Digital Spatial Profiling (DSP), a non-destructive method for high-plex spatial profiling of proteins and RNA, using oligonucleotide detection technologies with unlimited multiplexing capability. The key breakthroughs underlying DSP are threefold: (1) multiplexed readout of proteins/RNA using oligo-tags; (2) oligo-tags attached to affinity reagents (antibodies/RNA probes) through a photocleavable (PC) linker; (3) photocleaving light projected onto the tissue sample to release PC-oligos in any spatial pattern. Here we show precise analyte reproducibility, validation, and cellular resolution using DSP. We also demonstrate biological proof-of-concept using lymphoid, colorectal tumor, and autoimmune tissue as models to profile immune cell populations, stroma, and cancer cells to identify factors specific for the diseased microenvironment. DSP utilizes the unlimited multiplexing capability of modern genomic approaches, while simultaneously providing spatial context of protein and RNA to examine biological questions based on analyte location and distribution.