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Title: Multi-Scale LM/EM Neuronal Imaging from Brain to Synapse with a Tissue Clearing Method, ScaleSF

Takahiro Furuta, Kenta Yamauchi, Shinichiro Okamoto, Megumu Takahashi, Soichiro Kakuta, Yoko Ishida, Aya Takenaka, Atsushi Yoshida, Yasuo Uchiyama, Masato Koike, Kaoru Isa, Tadashi Isa, View ORCID ProfileHiroyuki Hioki
doi: https://doi.org/10.1101/2021.04.02.438164
Takahiro Furuta
1Department of Oral Anatomy and Neurobiology, Graduate School of Dentistry, Osaka University, Suita, Osaka 565-0871, Japan
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Kenta Yamauchi
2Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, Bunkyo-Ku, Tokyo 113-8421, Japan
3Advanced Research Institute for Health Sciences, Juntendo University, Bunkyo-Ku, Tokyo 113-8421, Japan
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Shinichiro Okamoto
2Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, Bunkyo-Ku, Tokyo 113-8421, Japan
3Advanced Research Institute for Health Sciences, Juntendo University, Bunkyo-Ku, Tokyo 113-8421, Japan
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Megumu Takahashi
2Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, Bunkyo-Ku, Tokyo 113-8421, Japan
4Department of Neuroscience, Graduate School of Medicine, Kyoto University, Kyoto, Kyoto 606-8501, Japan.
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Soichiro Kakuta
5Laboratory of Morphology and Image Analysis, Research Support Center, Juntendo University Graduate School of Medicine, Bunkyo-Ku, Tokyo 113-8421, Japan.
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Yoko Ishida
2Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, Bunkyo-Ku, Tokyo 113-8421, Japan
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Aya Takenaka
1Department of Oral Anatomy and Neurobiology, Graduate School of Dentistry, Osaka University, Suita, Osaka 565-0871, Japan
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Atsushi Yoshida
1Department of Oral Anatomy and Neurobiology, Graduate School of Dentistry, Osaka University, Suita, Osaka 565-0871, Japan
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Yasuo Uchiyama
6Department of Cellular and Molecular Neuropathology, Juntendo University Graduate School of Medicine, Bunkyo-Ku, Tokyo 113-8421, Japan.
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Masato Koike
2Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, Bunkyo-Ku, Tokyo 113-8421, Japan
3Advanced Research Institute for Health Sciences, Juntendo University, Bunkyo-Ku, Tokyo 113-8421, Japan
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Kaoru Isa
4Department of Neuroscience, Graduate School of Medicine, Kyoto University, Kyoto, Kyoto 606-8501, Japan.
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Tadashi Isa
4Department of Neuroscience, Graduate School of Medicine, Kyoto University, Kyoto, Kyoto 606-8501, Japan.
7Institute for the Advanced Study of Human Biology (WPI-ASHBi), Kyoto University, Kyoto, Kyoto 606-8501, Japan.
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Hiroyuki Hioki
2Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, Bunkyo-Ku, Tokyo 113-8421, Japan
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  • ORCID record for Hiroyuki Hioki
  • For correspondence: hioki-ns@umin.ac.jp
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Abstract

The mammalian brain is organized over sizes that span several orders of magnitude, from synapses to the entire brain. Thus, a technique to visualize neural circuits across multiple spatial scales (multi-scale neuronal imaging) is vital for deciphering brain-wide connectivity. Here, we developed this technique by coupling successive light microscope/electron microscope (LM/EM) imaging with an ultrastructurally-preserved tissue clearing method, ScaleSF. Our multi-scale neuronal imaging incorporates 1) brain-wide macroscopic observation, 2) mesoscopic circuit mapping, 3) microscopic subcellular imaging, and 4) EM imaging of nanoscopic structures, allowing seamless integration of structural information from the brain to synapses. We applied the technique to three neural circuits of two different species, mouse striatofugal, mouse callosal, and marmoset corticostriatal projection systems, and succeeded in the simultaneous interrogation of their circuit structure and synaptic connectivity in a targeted way. Our multi-scale neuronal imaging will significantly advance the understanding of brain-wide connectivity by expanding the scales of objects.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted April 02, 2021.
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Title: Multi-Scale LM/EM Neuronal Imaging from Brain to Synapse with a Tissue Clearing Method, ScaleSF
Takahiro Furuta, Kenta Yamauchi, Shinichiro Okamoto, Megumu Takahashi, Soichiro Kakuta, Yoko Ishida, Aya Takenaka, Atsushi Yoshida, Yasuo Uchiyama, Masato Koike, Kaoru Isa, Tadashi Isa, Hiroyuki Hioki
bioRxiv 2021.04.02.438164; doi: https://doi.org/10.1101/2021.04.02.438164
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Title: Multi-Scale LM/EM Neuronal Imaging from Brain to Synapse with a Tissue Clearing Method, ScaleSF
Takahiro Furuta, Kenta Yamauchi, Shinichiro Okamoto, Megumu Takahashi, Soichiro Kakuta, Yoko Ishida, Aya Takenaka, Atsushi Yoshida, Yasuo Uchiyama, Masato Koike, Kaoru Isa, Tadashi Isa, Hiroyuki Hioki
bioRxiv 2021.04.02.438164; doi: https://doi.org/10.1101/2021.04.02.438164

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