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SCRINSHOT enables spatial mapping of cell states in tissue sections with single-cell resolution.

ABSTRACT: Changes in cell identities and positions underlie tissue development and disease progression. Although single-cell mRNA sequencing (scRNA-Seq) methods rapidly generate extensive lists of cell states, spatially resolved single-cell mapping presents a challenging task. We developed SCRINSHOT (Single-Cell Resolution IN Situ Hybridization On Tissues), a sensitive, multiplex RNA mapping approach. Direct hybridization of padlock probes on mRNA is followed by circularization with SplintR ligase and rolling circle amplification (RCA) of the hybridized padlock probes. Sequential detection of RCA-products using fluorophore-labeled oligonucleotides profiles thousands of cells in tissue sections. We evaluated SCRINSHOT specificity and sensitivity on murine and human organs. SCRINSHOT quantification of marker gene expression shows high correlation with published scRNA-Seq data over a broad range of gene expression levels. We demonstrate the utility of SCRINSHOT by mapping the locations of abundant and rare cell types along the murine airways. The amenability, multiplexity, and quantitative qualities of SCRINSHOT facilitate single-cell mRNA profiling of cell-state alterations in tissues under a variety of native and experimental conditions.

SUBMITTER: Sountoulidis A

PROVIDER: S-EPMC7717588 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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SCRINSHOT enables spatial mapping of cell states in tissue sections with single-cell resolution.

Sountoulidis Alexandros A Liontos Andreas A Nguyen Hong Phuong HP Firsova Alexandra B AB Fysikopoulos Athanasios A Qian Xiaoyan X Seeger Werner W Sundström Erik E Nilsson Mats M Samakovlis Christos C

PLoS biology 20201120 11

Changes in cell identities and positions underlie tissue development and disease progression. Although single-cell mRNA sequencing (scRNA-Seq) methods rapidly generate extensive lists of cell states, spatially resolved single-cell mapping presents a challenging task. We developed SCRINSHOT (Single-Cell Resolution IN Situ Hybridization On Tissues), a sensitive, multiplex RNA mapping approach. Direct hybridization of padlock probes on mRNA is followed by circularization with SplintR ligase and rol ...[more]

PMID: 33216742

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Dataset Information

SCRINSHOT enables spatial mapping of cell states in tissue sections with single-cell resolution.

Publications

SCRINSHOT enables spatial mapping of cell states in tissue sections with single-cell resolution.

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