Journal
NATURE COMMUNICATIONS
Volume 11, Issue 1, Pages -Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41467-020-15906-5
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Funding
- PRESTO from JST [JPMJPR15F4]
- JSPS KAKENHI [15H05650, 19H03413, 18H02105, 17H06328, 17H05688, 18H04543, 18K19419, 18K19373, 16H02277, 19J13188, 18H05270]
- Takeda Science Foundation
- Japan Foundation for Applied Enzymology
- AMED-CREST (AMED/MEXT)
- CREST (JST/MEXT)
- Brain/MINDS (AMED/MEXT)
- Basic Science and Platform Technology Program for Innovative Biological Medicine (AMED/MEXT)
- Science and Technology Platform Program for Advanced Biological Medicine (AMED/MEXT)
- Collaborative Research Project of the Brain Research Institute, Niigata University
- Human Frontier Science Program
- JST, Japan [JPMJER1202]
- RIKEN BRC through the National BioResource Project of the MEXT/AMED, Japan [RBRC06344]
- Grants-in-Aid for Scientific Research [18K19419, 18K19373, 19H03413, 19J13188, 18H05270, 18H02105, 17H06328, 18H04543, 17H05688, 15H05650] Funding Source: KAKEN
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Whole-organ/body three-dimensional (3D) staining and imaging have been enduring challenges in histology. By dissecting the complex physicochemical environment of the staining system, we developed a highly optimized 3D staining imaging pipeline based on CUBIC. Based on our precise characterization of biological tissues as an electrolyte gel, we experimentally evaluated broad 3D staining conditions by using an artificial tissue-mimicking material. The combination of optimized conditions allows a bottom-up design of a superior 3D staining protocol that can uniformly label whole adult mouse brains, an adult marmoset brain hemisphere, an similar to 1 cm(3) tissue block of a postmortem adult human cerebellum, and an entire infant marmoset body with dozens of antibodies and cell-impermeant nuclear stains. The whole-organ 3D images collected by light-sheet microscopy are used for computational analyses and whole-organ comparison analysis between species. This pipeline, named CUBIC-HistoVIsion, thus offers advanced opportunities for organ- and organism-scale histological analysis of multicellular systems. Tissue clearing has revolutionised histology, but limited penetration of antibodies and stains into thick tissue segments is still a bottleneck. Here, the authors characterise optically cleared tissue as an electrolyte gel and apply this knowledge to stain the entirety of thick tissue samples.
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