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2023 Fiscal Year Final Research Report

Application of RAP tissue clearing protocol for immunofluorescent multiplex staining of whole mount specimen

Research Project

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Project/Area Number 21K12258
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 63030:Chemical substance influence on environment-related
Research InstitutionKanazawa Medical University

Principal Investigator

SAKATA-HAGA Hiromi  金沢医科大学, 医学部, 准教授 (50294666)

Co-Investigator(Kenkyū-buntansha) 八田 稔久  金沢医科大学, 医学部, 教授 (20238025)
Project Period (FY) 2021-04-01 – 2024-03-31
Keywords組織透明化 / 免疫染色 / マウス / 多重染色
Outline of Final Research Achievements

We have previously reported a rapid bone staining procedure (RAP-B) which
based on a rapid and non-destructive tissue clearing system (RAP). One of the notable advantages of RAP is that histological structure is preserved even after the tissue is highly transparent in whole-body bone-stained specimens of larger specimens than ever before. To take advantage of this feature, we have applied it for immunohistochemical analysis (RAP-IHC) and in situ hybridization (RAP-WISH) in whole-mount specimens of muse fetuses and thick-sliced tissue sections of various organs derived from adult rodents. In the present study, we examined the application of RAP tissue clearing protocol for fluorescence multiplex immunohistochemical staining using tyramide signal amplification (TSA) in whole-mount specimens of mouse

Free Research Field

先天異常学 発生毒性学

Academic Significance and Societal Importance of the Research Achievements

近年、組織透明化技術は、神経組織内部のネットワークの解析や、病理組織標本の解析などに広く使用され、無くてはならない技術となっている。我々の考案したRAPはアルカリ溶液であるにも関わらず、加温や長時間の反応を行っても軟部組織の崩壊が起こらず、標本の形態や骨格の原位置が保たれるという優れた特徴を持つ。他の組織透明化法で使われる試薬より安価、簡便、迅速などの点で優位であることが特徴である。また、研究は免疫染色等を行った後の標本を透徹する高屈折率マウント液は蛍光標識した標本にも使用可能であるうえ、安価な試薬の組み合わせで調整が可能であるうえ、水溶性であり、大変扱いやすい溶液となっており汎用性が高い。

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Published: 2025-01-30  

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