Histological and Morphological Characterization of Developing Dermal Lymphatic Vessels
Histological and Morphological Characterization of Developing Dermal Lymphatic Vessels
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DOI：
10.1007/978-1-4939-8712-2_2
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Harvey, Natasha L.
中科院分区：
文献类型：
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作者：
Betterman, Kelly L.;Harvey, Natasha L.
三维可视化淋巴管系统的能力彻底改变了我们对形态发生机制的理解，这对于发育过程中构建淋巴管网络非常重要。通常采用两种互补的方法来评估对小鼠胚胎中真皮淋巴管系统发育重要的基因和信号通路的功能。第一个是胚胎皮肤的整体免疫染色，以分析二维和三维的真皮淋巴管网络模式和形态。第二种是薄组织切片的免疫染色，以检查二维环境中离散淋巴管内皮细胞内的淋巴管身份、管腔形成和蛋白质定位。在这里，我们提出了胚胎背部皮肤和薄组织冷冻切片的多色免疫荧光免疫染色的详细方案。这些方法中的每一种都会生成真皮淋巴管系统的高分辨率图像，产生对发育中的小鼠胚胎中淋巴管表型的深入表征不可或缺的信息。
The capacity to visualize the lymphatic vasculature in three-dimensions has revolutionized our understanding of the morphogenetic mechanisms important for constructing the lymphatic vascular network during development. Two complementary approaches are commonly employed to assess the function of genes and signaling pathways important for development of the dermal lymphatic vasculature in the mouse embryo. The first of these is whole-mount immunostaining of embryonic skin to analyze dermal lymphatic vessel network patterning and morphology in two and three dimensions. The second is immunostaining of thin tissue sections to examine lymphatic vessel identity, lumen formation and protein localization within discrete lymphatic endothelial cells in a two-dimensional setting. Here we present detailed protocols for multicolor immunofluorescent immunostaining of embryonic dorsal skin and thin tissue cryosections. Each of these methods generates high-resolution images of the dermal lymphatic vasculature, yielding information integral to in-depth characterization of lymphatic vessel phenotypes in the developing mouse embryo.