Spatial analysis of tissue immunity and vascularity by light sheet fluorescence microscopy.
Spatial analysis of tissue immunity and vascularity by light sheet fluorescence microscopy.
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DOI：
10.1038/s41596-023-00941-5
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发表时间：
2024-01
期刊：
影响因子：
14.8
通讯作者：
Duo Zhang;Abigail H. Cleveland;Elisavet Krimitza;Katherine Han;Chenlong Yi;Andrea L Stout;Wei Zou;Jay F Dorsey;Yanqing Gong;Yi Fan
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文献类型：
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作者：
Duo Zhang;Abigail H. Cleveland;Elisavet Krimitza;Katherine Han;Chenlong Yi;Andrea L Stout;Wei Zou;Jay F Dorsey;Yanqing Gong;Yi Fan
癌症和心血管疾病的发病机制受到组织微环境的时空调节。包括免疫细胞、脉管系统和组织缺氧在内的微环境成分的多重可视化提供了疾病进展和治疗抗性的关键信息，这通常受到大体积组织中成像深度和分辨率的限制。为此，光片荧光显微镜，组织澄清和免疫染色后，可以产生三维高分辨率图像在整个器官水平。在这里，我们提供了一个详细的描述光片荧光显微镜成像分析的免疫细胞组成，血管化，组织灌注和缺氧在小鼠正常的大脑和心脏，以及脑肿瘤。我们描述了一个程序可视化组织血管化，灌注和缺氧与转基因血管标记系统。我们提供组织收集、组织半透明化和免疫染色的程序。我们进一步描述了用于分析组织免疫和血管分布的标准方法。我们预计，这种方法将有助于空间说明的结构和功能的组织微环境成分在癌症和心血管疾病。该程序需要1-2周，可以由具有一般分子生物学专业知识的用户执行。
The pathogenesis of cancer and cardiovascular diseases is subjected to spatiotemporal regulation by the tissue microenvironment. Multiplex visualization of the microenvironmental components, including immune cells, vasculature and tissue hypoxia, provides critical information underlying the disease progression and therapy resistance, which is often limited by imaging depth and resolution in large-volume tissues. To this end, light sheet fluorescence microscopy, following tissue clarification and immunostaining, may generate three-dimensional high-resolution images at a whole-organ level. Here we provide a detailed description of light sheet fluorescence microscopy imaging analysis of immune cell composition, vascularization, tissue perfusion and hypoxia in mouse normal brains and hearts, as well as brain tumors. We describe a procedure for visualizing tissue vascularization, perfusion and hypoxia with a transgenic vascular labeling system. We provide the procedures for tissue collection, tissue semi-clearing and immunostaining. We further describe standard methods for analyzing tissue immunity and vascularity. We anticipate that this method will facilitate the spatial illustration of structure and function of the tissue microenvironmental components in cancer and cardiovascular diseases. The procedure requires 1–2 weeks and can be performed by users with expertise in general molecular biology.