Versatile whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissues
Versatile whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissues
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DOI：
10.1038/s41467-020-15906-5
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发表时间：
2020-04-27
影响因子：
16.6
通讯作者：
Ueda, Hiroki R.
中科院分区：
文献类型：
--
作者：
Susaki, Etsuo A.;Shimizu, Chika;Ueda, Hiroki R.
全器官/身体三维（3D）染色和成像一直是组织学中的持久挑战。通过解剖染色系统复杂的物理化学环境，我们开发了一个高度优化的基于CUBIC的3D染色成像流水线。基于我们对生物组织作为电解质凝胶的精确表征，我们通过使用人工组织模拟材料实验评估了广泛的3D染色条件。优化条件的组合允许自下而上设计上级3D染色方案，该方案可以用数十种抗体和细胞不渗透的核染色剂均匀地标记整个成年小鼠脑、成年绒猴脑半球、死后成年人小脑的类似于1 cm（3）的组织块和整个婴儿绒猴体。光片显微镜收集的全器官3D图像用于计算分析和物种间的全器官比较分析。因此，这条名为CUBIC-HistoVIsion的管道为多细胞系统的器官和生物体规模的组织学分析提供了先进的机会。组织清除已经彻底改变了组织学，但抗体和染色剂对厚组织段的有限渗透仍然是一个瓶颈。在这里，作者将光学透明的组织作为电解质凝胶，并应用这一知识对整个厚组织样本进行染色。
Whole-organ/body three-dimensional (3D) staining and imaging have been enduring challenges in histology. By dissecting the complex physicochemical environment of the staining system, we developed a highly optimized 3D staining imaging pipeline based on CUBIC. Based on our precise characterization of biological tissues as an electrolyte gel, we experimentally evaluated broad 3D staining conditions by using an artificial tissue-mimicking material. The combination of optimized conditions allows a bottom-up design of a superior 3D staining protocol that can uniformly label whole adult mouse brains, an adult marmoset brain hemisphere, an similar to 1 cm(3) tissue block of a postmortem adult human cerebellum, and an entire infant marmoset body with dozens of antibodies and cell-impermeant nuclear stains. The whole-organ 3D images collected by light-sheet microscopy are used for computational analyses and whole-organ comparison analysis between species. This pipeline, named CUBIC-HistoVIsion, thus offers advanced opportunities for organ- and organism-scale histological analysis of multicellular systems. Tissue clearing has revolutionised histology, but limited penetration of antibodies and stains into thick tissue segments is still a bottleneck. Here, the authors characterise optically cleared tissue as an electrolyte gel and apply this knowledge to stain the entirety of thick tissue samples.