完整组织的高分辨率三维荧光成像

Danny El-Nachef, A. Martinson, Xiulan Yang, C. Murry, W., R. Maclellan
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引用次数: 0

摘要

荧光标记组织的组织学分析是了解原位分子组织的关键工具。然而,在人体器官解剖的背景下,评估大细胞内的分子结构一直具有挑战性,因为它需要染色试剂和光线深入不透明组织,同时也符合高分辨率物镜的空间限制。这篇方法学文章描述了亚微米分辨率人类和啮齿动物组织三维成像的优化样品制备,产生成像深度(>100 μ m)和分辨率(100 μ m厚)。这种方法可以很容易地被任何进行常规组织学的实验室采用,从组织准备开始到3D图像需要3天。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
High-resolution 3D fluorescent imaging of intact tissues
Histological analysis of fluorescently labeled tissues has been a critical tool to understand molecular organization in situ. However, assessing molecular structures within large cells and in the context of human organ anatomy has been challenging because it requires penetration of staining reagents and light deep into opaque tissues, while also conforming to the spatial constraints of high-resolution objective lenses. This methodology article describes optimized sample preparation for sub-micron resolution 3D imaging in human and rodent tissues, yielding imaging depth (>100 µm) and resolution (<0.012 µm3 voxel size) that has previously been limited to whole-mount in vitro organoid systems, embryos, and small model organisms. Confocal images of adult human and rodent organs, including heart, kidney, and liver, were generated for several chemical and antibody stains in cleared tissue sections >100 µm thick. This method can be readily adopted by any lab performing routine histology and takes 3 days from the start of tissue preparation to 3D images.
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