Versatile whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissues

Etsuo A. Susaki (), Chika Shimizu, Akihiro Kuno, Kazuki Tainaka, Xiang Li, Kengo Nishi, Ken Morishima, Hiroaki Ono, Koji L. Ode, Yuki Saeki, Kazunari Miyamichi, Kaoru Isa, Chihiro Yokoyama, Hiroki Kitaura, Masako Ikemura, Tetsuo Ushiku, Yoshihiro Shimizu, Takashi Saito, Takaomi C. Saido, Masashi Fukayama, Hirotaka Onoe, Kazushige Touhara, Tadashi Isa, Akiyoshi Kakita, Mitsuhiro Shibayama and Hiroki R. Ueda ()
Additional contact information
Etsuo A. Susaki: The University of Tokyo
Chika Shimizu: Laboratory for Synthetic Biology, RIKEN Center for Biosystems Dynamics Research
Akihiro Kuno: The University of Tokyo
Kazuki Tainaka: Niigata University
Xiang Li: The University of Tokyo
Kengo Nishi: The University of Tokyo
Ken Morishima: The University of Tokyo
Hiroaki Ono: The University of Tokyo
Koji L. Ode: The University of Tokyo
Yuki Saeki: The University of Tokyo
Kazunari Miyamichi: The University of Tokyo
Kaoru Isa: Kyoto University, Yoshida-konoe-cho
Chihiro Yokoyama: Laboratory for Brain Connectomics Imaging, RIKEN Center for Biosystems Dynamics Research, 6-7-3 Minatojima-minamimachi, Chuo-ku
Hiroki Kitaura: Brain Research Institute, Niigata University
Masako Ikemura: The University of Tokyo
Tetsuo Ushiku: The University of Tokyo
Yoshihiro Shimizu: Laboratory for Cell-Free Protein Synthesis, RIKEN Center for Biosystems Dynamics Research, 6-2-3, Furuedai
Takashi Saito: Laboratory for Proteolytic Neuroscience, RIKEN Center for Brain Science
Takaomi C. Saido: Laboratory for Proteolytic Neuroscience, RIKEN Center for Brain Science
Masashi Fukayama: The University of Tokyo
Hirotaka Onoe: Kyoto University
Kazushige Touhara: The University of Tokyo
Tadashi Isa: Kyoto University, Yoshida-konoe-cho
Akiyoshi Kakita: Brain Research Institute, Niigata University
Mitsuhiro Shibayama: The University of Tokyo
Hiroki R. Ueda: The University of Tokyo

Nature Communications, 2020, vol. 11, issue 1, 1-22

Abstract: Abstract 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.

Date: 2020
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DOI: 10.1038/s41467-020-15906-5

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