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

被引：143

作者：

Susaki, Etsuo A. ^{[1
,2
]}

Shimizu, Chika ^{[2
]}

Kuno, Akihiro ^{[1
,3
]}

Tainaka, Kazuki ^{[4
]}

Li, Xiang ^{[5
]}

Nishi, Kengo ^{[5
]}

Morishima, Ken ^{[5
]}

Ono, Hiroaki ^{[1
]}

Ode, Koji L. ^{[1
,2
]}

Saeki, Yuki ^{[6
]}

Miyamichi, Kazunari ^{[7
,8
]}

Isa, Kaoru ^{[9
,10
,11
]}

Yokoyama, Chihiro ^{[12
]}

Kitaura, Hiroki ^{[13
]}

Ikemura, Masako ^{[14
]}

Ushiku, Tetsuo ^{[14
]}

Shimizu, Yoshihiro ^{[15
]}

Saito, Takashi ^{[16
,17
]}

Saido, Takaomi C. ^{[16
]}

Fukayama, Masashi ^{[14
]}

Onoe, Hirotaka ^{[18
]}

Touhara, Kazushige ^{[7
,8
,19
]}

Isa, Tadashi ^{[9
,10
,11
]}

Kakita, Akiyoshi ^{[13
]}

Shibayama, Mitsuhiro ^{[5
]}

Ueda, Hiroki R. ^{[1
,2
]}

机构：

[1] Univ Tokyo, Grad Sch Med, Dept Syst Pharmacol, Bunkyo Ku, 7-3-1 Hongo, Tokyo 1130033, Japan

[2] RIKEN, Lab Synthet Biol, Ctr Biosyst Dynam Res, 1-3 Yamadaoka, Suita, Osaka 5655241, Japan

[3] Univ Tsukuba, Dept Anat & Embryol, Fac Med, 1-1-1 Tennodai, Tsukuba, Ibaraki 3058575, Japan

[4] Niigata Univ, Brain Res Inst, Dept Syst Pathol Neurol Disorders, Chuo Ku, 1-757 Asahimachidori, Niigata 9518585, Japan

[5] Univ Tokyo, Inst Solid State Phys, Neutron Sci Lab, 5-1-5 Kashiwanoha, Kashiwa, Chiba 2778581, Japan

[6] Univ Tokyo, Fac Med, Bunkyo Ku, 7-3-1 Hongo, Tokyo 1130033, Japan

[7] Univ Tokyo, Grad Sch Agr & Life Sci, Dept Appl Biol Chem, Bunkyo Ku, 1-1-1 Yayoi, Tokyo 1138657, Japan

[8] Univ Tokyo, ERATO Touhara Chemosensory Signal Project, Japan Sci & Technol Agcy, Bunkyo Ku, 1-1-1 Yayoi, Tokyo 1138657, Japan

[9] Kyoto Univ, Grad Sch Med, Dept Neurosci, Sakyo Ku, Yoshida Konoe Cho, Kyoto 6068501, Japan

[10] Kyoto Univ, Fac Med, Sakyo Ku, Yoshida Konoe Cho, Kyoto 6068501, Japan

[11] Kyoto Univ, Inst Adv Study Human Biol WPI ASHBi, Kyoto 6068501, Japan

[12] RIKEN, Lab Brain Connect Imaging, Ctr Biosyst Dynam Res, Chuo Ku, 6-7-3 Minatojima Minamimachi, Kobe, Hyogo 6500047, Japan

[13] Niigata Univ, Brain Res Inst, Dept Pathol, Chuo Ku, 1-757 Asahimachidori, Niigata 9518585, Japan

[14] Univ Tokyo, Grad Sch Med, Dept Pathol, Bunkyo Ku, 7-3-1 Hongo, Tokyo 1130033, Japan

[15] RIKEN, Lab Cell Free Prot Synth, Ctr Biosyst Dynam Res, 6-2-3 Furuedai, Suita, Osaka 5650874, Japan

[16] RIKEN, Lab Proteolyt Neurosci, Ctr Brain Sci, 2-1 Hirosawa, Wako, Saitama 3510198, Japan

[17] Nagoya City Univ, Grad Sch Med Sci, Dept Neurocognit Sci,Inst Brain Sci, Mizuho Ku, 1 Kawasumi,Mizuho Cho, Nagoya, Aichi 4678601, Japan

[18] Kyoto Univ, Grad Sch Med, Human Brain Res Ctr, Sakyo Ku, 54 Shogoin Kawahara Cho, Kyoto 6068507, Japan

[19] Univ Tokyo, Int Res Ctr Neurointelligence WPI IRCN, UTIAS, Tokyo 1130033, Japan

来源：

NATURE COMMUNICATIONS | 2020年 / 11卷 / 01期

关键词：

SINGLE-CELL RESOLUTION; EARLY BRAIN-INJURY; BODY; MICROSCOPY; CIRCUITS; FRACTALS; FEATURES; NEURONS; IDISCO;

D O I：

10.1038/s41467-020-15906-5

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：22

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