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Quick visualization of neurons in brain tissues using an optical clearing technique

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Abstract

Neurons are classified into several morphological types according to the locations of their somata and the branching patterns of their axons and dendrites. Recent studies suggest that these morphological features are related to their physiological properties, including firing characteristics, responses to neuromodulators, and wiring patterns. Therefore, rapid morphological identification of electrophysiologically recorded neurons promises to advance our understanding of neuronal circuits. One of the most common anatomical cell identification methods is neuronal reconstruction with biocytin delivered through whole-cell patch-clamp pipettes. However, conventional reconstruction methods usually take longer than 24 h and limit the throughput of electrophysiological experiments. Here, we developed a quick, simple cell reconstruction method by optimizing the tissue clearing protocol ScaleSQ. We found that adding 200 mM NaCl almost entirely prevented tissue swelling without compromising optical clearing ability. This solution, termed IsoScaleSQ, allowed us to increase the transparency of the gray matter of 500-µm-thick slices within 30 min, meaning that the total time required to reconstruct whole-cell recorded neurons was reduced to 1 h. This novel method will improve the efficacy and effectiveness of electrophysiological experiments linked to cell morphology.

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Acknowledgements

This work was supported by JST ERATO (JPMJER1801), JSPS Grants-in-Aid for Scientific Research (18H05525), and the Human Frontier Science Program (RGP0019/2016). This work was conducted partly as a program at the International Research Center for Neurointelligence (WPI-IRCN) of The University of Tokyo Institutes for Advanced Study at The University of Tokyo.

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Authors and Affiliations

  1. Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan

    Yu Sato, Takeyuki Miyawaki, Ayako Ouchi, Asako Noguchi & Yuji Ikegaya

  2. Department of Morphological Neuroscience, Graduate School of Medicine, Gifu University, 1-1 Yanagido, Gifu City, Gifu, 501-1194, Japan

    Shun Yamaguchi

  3. Center for Highly Advanced Integration of Nano and Life Sciences, Gifu University, 1-1 Yanagido, Gifu City, Gifu, 501-1194, Japan

    Shun Yamaguchi

  4. Center for Information and Neural Networks, National Institute of Information and Communications Technology, Suita City, Osaka, 565-0871, Japan

    Yuji Ikegaya

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  1. Yu Sato
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  2. Takeyuki Miyawaki
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  3. Ayako Ouchi
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  4. Asako Noguchi
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  5. Shun Yamaguchi
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  6. Yuji Ikegaya
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Correspondence to Takeyuki Miyawaki.

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Sato, Y., Miyawaki, T., Ouchi, A. et al. Quick visualization of neurons in brain tissues using an optical clearing technique. Anat Sci Int 94, 199–208 (2019). https://doi.org/10.1007/s12565-018-00473-z

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