Whole-brain spatial transcriptional analysis at cellular resolution
Recent advances in RNA analysis have deepened our understanding of cellular states in biological tissues. However, a substantial gap remains in integrating RNA expression data with spatial context across organs, primarily owing to the challenges associated with RNA detection within intact tissue volumes. Here, we developed Tris buffer-mediated retention of in situ hybridization chain reaction signal in cleared organs (TRISCO), an effective tissue-clearing method designed for whole-brain spatial three-dimensional (3D) RNA imaging. TRISCO resolved several crucial issues, including the preservation of RNA integrity, achieving uniform RNA labeling, and enhancing tissue transparency. We tested TRISCO using a broad range of cell-identity markers, noncoding and activity-dependent RNAs, within diverse organs of varying sizes and species. TRISCO thus emerges as a powerful tool for single-cell, whole-brain, 3D imaging that enables comprehensive transcriptional spatial analysis across the entire brain.
Authors
- Jacob Lercke Skytte (ORCID: https://orcid.org/0000-0003-0313-2124)
- Casper G. Salinas (ORCID: https://orcid.org/0000-0003-3485-8331)
- Johan Hartman (ORCID: https://orcid.org/0000-0002-6500-8527)
- Mikael Adner (ORCID: https://orcid.org/0000-0003-1634-4759)
- Tsutomu Tanabe (ORCID: https://orcid.org/0000-0003-3000-9105)
- Daisuke H. Tanaka (ORCID: https://orcid.org/0000-0002-1435-709X)
- Keishiro Fukumoto (ORCID: https://orcid.org/0000-0002-5941-4000)
- Per Uhlén (ORCID: https://orcid.org/0000-0003-1446-1062)
- Naofumi Uesaka (ORCID: https://orcid.org/0000-0002-9744-944X)
- Ayako Miyakawa
- Shigeaki Kanatani (ORCID: https://orcid.org/0000-0003-2226-4288)
- Yujiao Xiang
- Gonçalo Castelo‐Branco (ORCID: https://orcid.org/0000-0003-2247-9393)
- Urmas Roostalu (ORCID: https://orcid.org/0000-0002-7849-2055)
- Songbai Zhang (ORCID: https://orcid.org/0000-0002-2033-8389)
- Razieh Karamzadeh
- Leslie A. Kirby (ORCID: https://orcid.org/0000-0003-4467-2661)
- Kimiharu Takamatsu (ORCID: https://orcid.org/0000-0002-3325-6606)
- Ilse Eidhof (ORCID: https://orcid.org/0000-0002-1567-3078)
- Zoe West (ORCID: https://orcid.org/0000-0002-1238-6444)
- Jacob Hecksher‐Sørensen (ORCID: https://orcid.org/0000-0002-5516-6177)
- Danai Vougesi Nikou
- Judith C. Kreutzmann (ORCID: https://orcid.org/0000-0003-0180-3449)
- Lea Lydolph Larsen
- Dagmara Kaczyńska
- Erik Sundström (ORCID: https://orcid.org/0000-0003-2931-8015)
- Xiaofei Li (ORCID: https://orcid.org/0000-0002-9991-7534)
- Yue Li (ORCID: https://orcid.org/0000-0003-0584-1119)
- A. G. Walton
Institutions
- Tokyo Medical and Dental University (JP)
- Karolinska University Hospital (SE)
- Keio University (JP)
- Karolinska Institutet (SE)
- Gubra (Denmark) (DK)
- Institute of Science Tokyo (JP)
- The University of Tokyo (JP)
Publication Details
- Journal
- Science
- Published
- 2024-11-21
- DOI
- https://doi.org/10.1126/science.adn9947
- Citations
- 40
- Primary Topic
- Single-cell and spatial transcriptomics
- Type
- article
- Field-Weighted Citation Impact
- 9.35