Versatile whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissues
| Autor: | Hiroki Kitaura, Tadashi Isa, Yoshihiro Shimizu, Yuki Saeki, Kazushige Touhara, Ken Morishima, Etsuo A. Susaki, Xiang Li, Kaoru Isa, Kazuki Tainaka, Masashi Fukayama, Takaomi C. Saido, Hiroki R. Ueda, Akihiro Kuno, Takashi Saito, Tetsuo Ushiku, Chika Shimizu, Mitsuhiro Shibayama, Hiroaki Ono, Akiyoshi Kakita, Chihiro Yokoyama, Kengo Nishi, Masako Ikemura, Hirotaka Onoe, Kazunari Miyamichi, Koji L. Ode |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Adult
Male 0301 basic medicine Fluorescence-lifetime imaging microscopy Science General Physics and Astronomy Electrolyte Stain Article Fluorescence imaging General Biochemistry Genetics and Molecular Biology Electrolytes Mice 03 medical and health sciences Imaging Three-Dimensional 0302 clinical medicine 3-D reconstruction Cerebellum biology.animal Microscopy Image Processing Computer-Assisted Animals Humans lcsh:Science Fluorescent Dyes Multidisciplinary biology Chemistry Optical Imaging Brain Marmoset Callithrix Histology General Chemistry Staining Mice Inbred C57BL 030104 developmental biology Animals Newborn Microscopy Fluorescence Organ body Female lcsh:Q 030217 neurology & neurosurgery Biomedical engineering |
| Zdroj: | Nature Communications, Vol 11, Iss 1, Pp 1-22 (2020) Nature Communications |
| ISSN: | 2041-1723 |
| Popis: | 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 ~1 cm3 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. |
| Databáze: | OpenAIRE |
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