2019
DOI: 10.1101/559021
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High multiplex, digital spatial profiling of proteins and RNA in fixed tissue using genomic detection methods

Abstract: 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… Show more

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Cited by 34 publications
(41 citation statements)
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“…However, correlation between DSP and alternative analysis methods has been described recently, indicating strong correlation with standard IHC techniques. 23,27 Indeed, when the mean CD3 and CD45 counts of the ROIs in the healthy BM sample were compared with DSP counts, good correlation was seen for both antigens ( Figure 1C).…”
Section: Resultsmentioning
confidence: 84%
See 1 more Smart Citation
“…However, correlation between DSP and alternative analysis methods has been described recently, indicating strong correlation with standard IHC techniques. 23,27 Indeed, when the mean CD3 and CD45 counts of the ROIs in the healthy BM sample were compared with DSP counts, good correlation was seen for both antigens ( Figure 1C).…”
Section: Resultsmentioning
confidence: 84%
“…Technologies laboratories under an early technology access programme, as described recently. 22,26,27 ROI selection and subsequent statistical and correlative analyses were undertaken in the Australian Cancer Research Foundation translational research laboratory. Briefly, a multiplexed cocktail of primary antibodies, each with a unique, UV-photocleavable indexing oligo (Table 1), mouse anti-human CD3E antibody clone UMAB54 (Origene Technologies, Inc., Rockville, MD) conjugated to AF647 using AF647 antibody labelling kit (Thermo Fisher Scientific, Waltham, MA), rabbit anti-human CD45 antibody clone D9M81 (Cell Signalling Technology, Inc., Danvers, MA) conjugated to AF594 using AF594 antibody labelling kit (Thermo Fisher Scientific) and Syto83 nucleic acid stain (Thermo Fisher Scientific), was applied to a slide-mounted BM trephine tissue section.…”
Section: Bm Trephine Sections Underwent Dsp At Nanostringmentioning
confidence: 99%
“…STvEA is complementary to emerging technologies for simultaneous highly-multiplexed spatial profiling of proteins and transcripts, such as Digital Spatial Profiling 40…”
Section: Discussionmentioning
confidence: 99%
“…Profiling every cell at single‐cell resolution using DSP may be impractical and cost‐prohibitive. Thus, a general advantage of imaging‐based methods compared to DSP is the ability to get multiplexed information on each cell in the tissue slice rather than high‐plex information of regions of interest as in DSP [91]. In other words, the DSP can only be visualized based on its ‘morphology kit’ of up to 3 markers, including cytokeratin and CD45.…”
Section: Brightfield‐based Mihc/ifmentioning
confidence: 99%
“…HALO (Indica Labs, Albuquerque, New Mexico, USA, ; https://www.indicalab.com/halo/) is an image analysis platform for quantitative tissue analysis in digital pathology, mainly used for analysis in oncology, toxicology, metabolism, neuroscience, and more [90‐94]. It reports the morphology and multiplexed expression data of each and every cell across the entire tissue sections.…”
Section: Brightfield‐based Mihc/ifmentioning
confidence: 99%