Multiplex, quantitative cellular analysis in large tissue volumes with clearing-enhanced 3D microscopy (Ce3D)
Multiplex, quantitative cellular analysis in large tissue volumes with clearing-enhanced 3D microscopy (Ce3D)
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DOI：
10.1073/pnas.1708981114
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发表时间：
2017-08-29
影响因子：
11.1
通讯作者：
Gerner, Michael Y.
中科院分区：
文献类型：
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作者：
Li, Weizhe;Germain, Ronald N.;Gerner, Michael Y.
器官稳态、细胞分化、信号传递和原位功能都依赖于复杂组织中细胞的空间组织。出于这个原因，在宏观组织结构的背景下，细胞定位，表型身份和功能状态的全面，高分辨率的映射对于更深入地理解不同的生物过程至关重要。在这里，我们报告了一种易于使用的方法，透明增强3D（Ce3D），它为大多数器官产生了极好的组织透明度，保留了细胞形态和蛋白质荧光，并且与基于抗体的免疫标记牢固兼容。这种增强的信号质量和能力，广泛的探针复用允许定量分析不同的，高度混杂的细胞群体在完整的Ce3D处理的组织通过3D组织细胞术。我们使用这种技术来展示淋巴和非淋巴器官中不同细胞类型的大体积，高分辨率显微镜，以及对淋巴组织中多个细胞群的组成和组织分布进行定量分析。结合组织细胞术，Ce3D提供了一个全面的战略，体积定量成像和分析，桥梁之间的差距差距传统的部分成像和解离为基础的技术。
Organ homeostasis, cellular differentiation, signal relay, and in situ function all depend on the spatial organization of cells in complex tissues. For this reason, comprehensive, high-resolution mapping of cell positioning, phenotypic identity, and functional state in the context of macroscale tissue structure is critical to a deeper understanding of diverse biological processes. Here we report an easy to use method, clearing-enhanced 3D (Ce3D), which generates excellent tissue transparency for most organs, preserves cellular morphology and protein fluorescence, and is robustly compatible with antibody-based immunolabeling. This enhanced signal quality and capacity for extensive probe multiplexing permits quantitative analysis of distinct, highly intermixed cell populations in intact Ce3D-treated tissues via 3D histo-cytometry. We use this technology to demonstrate large-volume, high-resolution microscopy of diverse cell types in lymphoid and nonlymphoid organs, as well as to perform quantitative analysis of the composition and tissue distribution of multiple cell populations in lymphoid tissues. Combined with histo-cytometry, Ce3D provides a comprehensive strategy for volumetric quantitative imaging and analysis that bridges the gap between conventional section imaging and disassociation-based techniques.