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Spatial transcriptome sequencing of FFPE tissues at the cellular level

Yang Liu, Archibald Enninful, Yanxiang Deng, View ORCID ProfileRong Fan
doi: https://doi.org/10.1101/2020.10.13.338475
Yang Liu
1Department of Biomedical Engineering, Yale University, New Haven, CT 06520, USA
2Yale Stem Cell Center and Yale Cancer Center, Yale School of Medicine, New Haven, CT 06520, USA
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Archibald Enninful
1Department of Biomedical Engineering, Yale University, New Haven, CT 06520, USA
2Yale Stem Cell Center and Yale Cancer Center, Yale School of Medicine, New Haven, CT 06520, USA
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Yanxiang Deng
1Department of Biomedical Engineering, Yale University, New Haven, CT 06520, USA
2Yale Stem Cell Center and Yale Cancer Center, Yale School of Medicine, New Haven, CT 06520, USA
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Rong Fan
1Department of Biomedical Engineering, Yale University, New Haven, CT 06520, USA
2Yale Stem Cell Center and Yale Cancer Center, Yale School of Medicine, New Haven, CT 06520, USA
3Human and Translational Immunology Program, Yale School of Medicine, New Haven, CT 06520, USA
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  • ORCID record for Rong Fan
  • For correspondence: rong.fan@yale.edu
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Abstract

Formalin-fixed paraffin-embedded (FFPE) tissues are the most abundant archivable specimens in clinical tissue banks, but unfortunately incompatible with single-cell level transcriptome sequencing due to RNA degradation in storage and RNA damage in extraction. We developed an in-tissue barcoding approach namely DBiT-seq for spatially revolved whole transcriptome sequencing at cellular level, which required no tissue dissociation or RNA exaction, thus potentially more suited for FFPE samples. Herein, we demonstrated spatial transcriptome sequencing of embryonic and adult mouse FFPE tissue sections at cellular level (25μm pixel size) with high coverage (>1,000 genes per pixel). Spatial transcriptome of a E10.5 mouse embryo identified all major anatomical features in the brain and abdominal region. Integration with singlecell RNA-seq data for cell type identification indicated that most tissue pixels were dominated by single-cell transcriptional phenotype. Spatial mapping of adult mouse aorta, atrium, and ventricle tissues identified the spatial distribution of different cell types. Spatial transcriptome sequencing of FFPE samples at cellular level may provide enormous opportunities in a wide range of biomedical research. It may allow us to revisit retrospectively the huge resource of clinical tissue specimens to study human disease mechanisms for the discovery of tissue biomarkers and therapeutic targets

Competing Interest Statement

R.F. is scientific founder and advisor of IsoPlexis Singleron Biotechnologies and AtlasXomics. The interests of R.F. were reviewed and managed by Yale University Provost Office in accordance with the University conflict of interest policies.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted October 14, 2020.
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Spatial transcriptome sequencing of FFPE tissues at the cellular level
Yang Liu, Archibald Enninful, Yanxiang Deng, Rong Fan
bioRxiv 2020.10.13.338475; doi: https://doi.org/10.1101/2020.10.13.338475
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Spatial transcriptome sequencing of FFPE tissues at the cellular level
Yang Liu, Archibald Enninful, Yanxiang Deng, Rong Fan
bioRxiv 2020.10.13.338475; doi: https://doi.org/10.1101/2020.10.13.338475

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