Simplified three-dimensional tissue clearing and incorporation of colorimetric phenotyping.
Simplified three-dimensional tissue clearing and incorporation of colorimetric phenotyping.
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DOI：
10.1038/srep30736
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发表时间：
2016-08-08
影响因子：
4.6
通讯作者：
Kulkarni RP
中科院分区：
文献类型：
--
作者：
Sung K;Ding Y;Ma J;Chen H;Huang V;Cheng M;Yang CF;Kim JT;Eguchi D;Di Carlo D;Hsiai TK;Nakano A;Kulkarni RP
组织清除方法有望提供精致的三维成像信息；然而，有必要简化方法的低资源设置和非荧光为基础的表型，使光显微镜成像模式。在这里，我们描述了简化的CLARITY方法（SCM）用于组织清理，保留感兴趣的表位。我们使用薄层显微镜对生成的组织进行成像，以生成整个组织和器官的快速3D重建。此外，为了使用光学显微镜方法进行清除和3D组织成像，我们开发了一种比色法，非荧光方法，用于特异性标记清除组织，该方法基于辣根过氧化物酶将二氨基联苯胺转化为有色不溶性产物。我们在这里描述的方法是便携式的，可以以低成本完成，并且可以允许清除组织的光显微成像，从而使组织清除和成像在各种各样的设置。
Tissue clearing methods promise to provide exquisite three-dimensional imaging information; however, there is a need for simplified methods for lower resource settings and for non-fluorescence based phenotyping to enable light microscopic imaging modalities. Here we describe the simplified CLARITY method (SCM) for tissue clearing that preserves epitopes of interest. We imaged the resulting tissues using light sheet microscopy to generate rapid 3D reconstructions of entire tissues and organs. In addition, to enable clearing and 3D tissue imaging with light microscopy methods, we developed a colorimetric, non-fluorescent method for specifically labeling cleared tissues based on horseradish peroxidase conversion of diaminobenzidine to a colored insoluble product. The methods we describe here are portable and can be accomplished at low cost, and can allow light microscopic imaging of cleared tissues, thus enabling tissue clearing and imaging in a wide variety of settings.