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

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  • Etsuo A. Susaki

    (The University of Tokyo
    Laboratory for Synthetic Biology, RIKEN Center for Biosystems Dynamics Research)

  • Chika Shimizu

    (Laboratory for Synthetic Biology, RIKEN Center for Biosystems Dynamics Research)

  • Akihiro Kuno

    (The University of Tokyo
    University of Tsukuba)

  • 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
    Laboratory for Synthetic Biology, RIKEN Center for Biosystems Dynamics Research)

  • Yuki Saeki

    (The University of Tokyo)

  • Kazunari Miyamichi

    (The University of Tokyo
    ERATO Touhara Chemosensory Signal Project, Japan Science and Technology Agency, The University of Tokyo)

  • Kaoru Isa

    (Kyoto University, Yoshida-konoe-cho
    Kyoto University)

  • 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
    Nagoya City University Graduate School of Medical Science, 1 Kawasumi, Mizuho-cho, Mizuho-ku)

  • 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
    ERATO Touhara Chemosensory Signal Project, Japan Science and Technology Agency, The University of Tokyo
    International Research Center for Neurointelligence (WPI-IRCN), UTIAS, The University of Tokyo)

  • Tadashi Isa

    (Kyoto University, Yoshida-konoe-cho
    Kyoto University)

  • Akiyoshi Kakita

    (Brain Research Institute, Niigata University)

  • Mitsuhiro Shibayama

    (The University of Tokyo)

  • Hiroki R. Ueda

    (The University of Tokyo
    Laboratory for Synthetic Biology, RIKEN Center for Biosystems Dynamics Research)

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.

Suggested Citation

  • 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 & , 2020. "Versatile whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissues," Nature Communications, Nature, vol. 11(1), pages 1-22, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-15906-5
    DOI: 10.1038/s41467-020-15906-5
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