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. 2022 Jun 28;14(13):3170. doi: 10.3390/cancers14133170

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Digital Spatial Profiling (DSP) workflow. Tissue sections are labeled with antibodies and/or in situ hybridization with mRNA probes, which are linked with UV-cleavable oligo-tags. Slides are labeled with fluorescence-conjugated antibodies to determine cell subsets and select regions of interests and masks for bulk cell subsets for directed UV-cleavage of the oligo-tags. The cleaved oligos are collected with a microcapillary and transferred to a 96-well plate. Next, the oligos are quantified by digital counting (nCounter) or next-generation sequencing. Differential expression of specific mRNA or proteins between ROIs and cell subsets are next analyzed (data analysis).