Bringing CLARITY to the human brain: visualization of Lewy pathology in three dimensions.
Bringing CLARITY to the human brain: visualization of Lewy pathology in three dimensions.
复制标题
DOI：
10.1111/nan.12293
复制
发表时间：
2016-10
影响因子：
5
通讯作者：
Gentleman SM
中科院分区：
文献类型：
--
作者：
Liu AK;Hurry ME;Ng OT;DeFelice J;Lai HM;Pearce RK;Wong GT;Chang RC;Gentleman SM
Clarity是一种新技术，它能够对免疫染色的组织进行三维可视化，以研究大脑中细胞和分子之间的电路和空间相互作用。在这项研究中，我们旨在比较啮齿类动物和大量人死后脑样本在透明度应用方面的方法学差异。此外，我们的目标是调查这项技术是否可以用于在帕金森氏病死后大脑中可视化路易病理。啮齿动物和人脑样本被澄清，并使用被动版本的Clarity技术进行免疫染色。然后将样品浸入不同的折射率匹配介质中，然后安装在共焦显微镜下进行可视化。我们发现，使用被动清晰度的组织清除速度因物种(人与啮齿动物)、大脑区域和固定程度(新鲜与福尔马林固定的组织)而不同。此外，使用特定的折射率匹配介质也有优势。我们已经应用这项技术，并成功地在三维可视化Meynert基底核内的路易体包涵体，以及中脑黑质纹状体纤维之间的单胺能纤维和路易氏病变之间的空间关系，而不需要对脑组织进行物理连续切片。对人体组织大样本的有效使用为详细的病理和形态研究开辟了许多潜在的途径。
CLARITY is a novel technique which enables three‐dimensional visualization of immunostained tissue for the study of circuitry and spatial interactions between cells and molecules in the brain. In this study, we aimed to compare methodological differences in the application of CLARITY between rodent and large human post mortem brain samples. In addition, we aimed to investigate if this technique could be used to visualize Lewy pathology in a post mortem Parkinson's brain. Rodent and human brain samples were clarified and immunostained using the passive version of the CLARITY technique. Samples were then immersed in different refractive index matching media before mounting and visualizing under a confocal microscope. We found that tissue clearing speed using passive CLARITY differs according to species (human vs. rodents), brain region and degree of fixation (fresh vs. formalin‐fixed tissues). Furthermore, there were advantages to using specific refractive index matching media. We have applied this technique and have successfully visualized Lewy body inclusions in three dimensions within the nucleus basalis of Meynert, and the spatial relationship between monoaminergic fibres and Lewy pathologies among nigrostriatal fibres in the midbrain without the need for physical serial sectioning of brain tissue. The effective use of CLARITY on large samples of human tissue opens up many potential avenues for detailed pathological and morphological studies.
登录
查看更多内容
影响因子：
--
作者：
Becker, Klaus;Jahrling, Nina;Dodt, Hans-Ulrich
通讯作者：
Dodt, Hans-Ulrich
影响因子：
6.4
作者：
Kanazawa, Toshiro;Uchihara, Toshiki;Mizusawa, Hidehiro
通讯作者：
Mizusawa, Hidehiro
影响因子：
3.7
作者：
Becker N;Jöst H;Ziegler U;Eiden M;Höper D;Emmerich P;Fichet-Calvet E;Ehichioya DU;Czajka C;Gabriel M;Hoffmann B;Beer M;Tenner-Racz K;Racz P;Günther S;Wink M;Bosch S;Konrad A;Pfeffer M;Groschup MH;Schmidt-Chanasit J
通讯作者：
Schmidt-Chanasit J
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
14.8
作者：
通讯作者：
--