Advanced CLARITY for rapid and high-resolution imaging of intact tissues.
Advanced CLARITY for rapid and high-resolution imaging of intact tissues.
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DOI：
10.1038/nprot.2014.123
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发表时间：
2014-07
期刊：
影响因子：
14.8
通讯作者：
中科院分区：
文献类型：
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CLARITY 是一种将完整的生物组织化学转化为水凝胶-组织混合物的方法，该混合物可以接受光和大分子标记的研究，同时保留精细结构和天然生物分子。这种从大型完整样本中获取信息的新兴方式既创造了新的机遇，也带来了新的挑战。在这里，我们描述了跨越 CLARITY 工作流程多个维度的下一代协议，从无需电泳仪器即可简单、可靠和高效去除脂质的新颖方法（被动 CLARITY），到优化物镜以及与光片光学器件（CLARITY 优化的光片显微镜或 COLM）的集成，以将澄清样品的数据收集速度加快几个数量级，同时保持或提高质量和分辨率。这些方法可以应用于大型组装生物系统（例如完整哺乳动物大脑）的结构和分子分析。
CLARITY is a method for chemical transformation of intact biological tissues into a hydrogel-tissue hybrid, which becomes amenable to interrogation with light and macromolecular labels while retaining fine structure and native biological molecules. This emerging accessibility of information from large intact samples has created both new opportunities and new challenges. Here we describe next-generation protocols spanning multiple dimensions of the CLARITY workflow, ranging from a novel approach to simple, reliable, and efficient lipid removal without electrophoretic instrumentation (passive CLARITY), to optimized objectives and integration with light-sheet optics (CLARITY-optimized light-sheet microscopy or COLM) for accelerating data collection from clarified samples by several orders of magnitude while maintaining or increasing quality and resolution. These methods may find application in the structural and molecular analysis of large assembled biological systems such as the intact mammalian brain.