| Project/Area Number |
18K11659
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 63030:Chemical substance influence on environment-related
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| Research Institution | Kanazawa Medical University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
八田 稔久 金沢医科大学, 医学部, 教授 (20238025)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | 骨染色 / 免疫染色 / マウス / コラーゲン誘発関節炎 / 組織透明化 / ３D画像解析 / ラット / 多重染色 / ３Ｄ画像解析 |
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| 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 system). We applied RAP-B not only to mouse and rat fetuses, but also to adult mice by combination with a hair removal procedure (RAP-B/HR). It was revealed that the RAP system was available to preparation of highly transparent bone-stained specimens of the mouse fetuses and adult mice. Observations by fluorescence zoom microscopy of these specimens delineated severe pathological bone feature such as joint deformities, narrowing of the joint fissure, bone fusion, as well as fine bone surface pathologies, such as bone erosion and osteophyte formation in the swollen joint area. RAP tissue clearing system also improved antibody permeation in both thick tissue sections and whole-mount specimens. Thus, we concluded that RAP tissue clearing system would be a powerful tool for immunohistochemical analysis in whole mount and thick-sliced specimens.
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| Academic Significance and Societal Importance of the Research Achievements |
近年、組織透明化技術は、骨格の観察だけでなく、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に応用されている。古典的な組織透明化はアルカリ溶液による軟部組織の脱色・透明化を基本としているが、この処置は時に過剰な組織破壊をもたらすことが問題となっていた。そのため、近年新しい組織透明化法が複数考案されている。我々の考案したRAP法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。近年開発された他の組織透明化法とは異なる組成のものであり、安価、簡便、迅速などの点で優位であることが特徴である。
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