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Anatomical Science International

, Volume 94, Issue 2, pp 199–208 | Cite as

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

  • Yu Sato
  • Takeyuki MiyawakiEmail author
  • Ayako Ouchi
  • Asako Noguchi
  • Shun Yamaguchi
  • Yuji Ikegaya
Original Article

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.

Keywords

Isotropic Optical clearing Patch-clamp recording Visualization ScaleSQ 

Notes

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.

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest to declare

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Copyright information

© Japanese Association of Anatomists 2019

Authors and Affiliations

  1. 1.Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical SciencesThe University of TokyoTokyoJapan
  2. 2.Department of Morphological Neuroscience, Graduate School of MedicineGifu UniversityGifu CityJapan
  3. 3.Center for Highly Advanced Integration of Nano and Life SciencesGifu UniversityGifu CityJapan
  4. 4.Center for Information and Neural NetworksNational Institute of Information and Communications TechnologySuita CityJapan

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