Sung Kevin, Ding Yichen, Ma Jianguo, Chen Harrison, Huang Vincent, Cheng Michelle, Yang Cindy F, Kim Jocelyn T, Eguchi Daniel, Di Carlo Dino, Hsiai Tzung K, Nakano Atsushi, Kulkarni Rajan P
Department of Bioengineering, University of California, Los Angeles, CA 90095, Los Angeles, USA.
Division of Cardiology, Department of Medicine, David Geffen School of Medicine at UCLA, CA 90095, Los Angeles, USA.
Sci Rep. 2016 Aug 8;6:30736. doi: 10.1038/srep30736.
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.
组织透明化方法有望提供精确的三维成像信息；然而，对于资源较少的环境以及基于非荧光的表型分析而言，需要简化方法以实现光学显微镜成像模式。在此，我们描述了一种用于组织透明化的简化CLARITY方法（SCM），该方法可保留感兴趣的表位。我们使用光片显微镜对所得组织进行成像，以生成整个组织和器官的快速三维重建。此外，为了能够通过光学显微镜方法进行透明化处理和三维组织成像，我们开发了一种比色、非荧光方法，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，对透明化组织进行特异性标记。我们在此描述的方法便于携带，成本低廉，能够对透明化组织进行光学显微镜成像，从而在各种环境中实现组织透明化和成像。
