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A Rapid Optical Clearing Protocol Using 2,2'-Thiodiethanol for Microscopic Observation of Fixed Mouse Brain

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Title: A Rapid Optical Clearing Protocol Using 2,2'-Thiodiethanol for Microscopic Observation of Fixed Mouse Brain
Authors: Aoyagi, Yuka Browse this author
Kawakami, Ryosuke Browse this author →KAKEN DB
Osanai, Hisayuki Browse this author
Hibi, Terumasa Browse this author →KAKEN DB
Nemoto, Tomomi Browse this author →KAKEN DB
Issue Date: 29-Jan-2015
Publisher: The Public Library of Science
Journal Title: Plos One
Volume: 10
Issue: 1
Start Page: e0116280
Publisher DOI: 10.1371/journal.pone.0116280
Abstract: Elucidation of neural circuit functions requires visualization of the fine structure of neurons in the inner regions of thick brain specimens. However, the tissue penetration depth of laser scanning microscopy is limited by light scattering and/or absorption by the tissue. Recently, several optical clearing reagents have been proposed for visualization in fixed specimens. However, they require complicated protocols or long treatment times. Here we report the effects of 2,2'-thiodiethanol (TDE) solutions as an optical clearing reagent for fixed mouse brains expressing a yellow fluorescent protein. Immersion of fixed brains in TDE solutions rapidly (within 30 min in the case of 400-mu m-thick fixed brain slices) increased their transparency and enhanced the penetration depth in both confocal and two-photon microscopy. In addition, we succeeded in visualizing dendritic spines along single dendrites at deep positions in fixed thick brain slices. These results suggest that our proposed protocol using TDE solution is a rapid and useful method for optical clearing of fixed specimens expressing fluorescent proteins.
Rights: http://creativecommons.org/licenses/by/4.0/
Type: article
URI: http://hdl.handle.net/2115/58426
Appears in Collections:電子科学研究所 (Research Institute for Electronic Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 根本 知己

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