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整片组织免疫染色显示淋巴管构筑
Architecture of the lymphatic vessel revealed with the whole-mount immunostaining
目的 探讨整片组织内淋巴管免疫染色，并与间接注射法和组织切片免疫染色法比较。方法 用普鲁士蓝间接注射新生儿肝被膜内淋巴管，观察肝浅淋巴管的构筑。通过火棉胶切片分析肝浅淋巴管的流向。取人大腿皮肤，作石蜡切片和冷冻切片，免疫染色后观察淋巴管的分布。取大鼠耳和背部的皮肤、膈和小肠，将整片组织作免疫染色，观察组织内淋巴管构筑。 结果 间接注射普鲁士蓝可显示淋巴管构筑和流向，但淋巴管染色缺乏特异性。组织切片免疫染色可显示淋巴管分布，然而不能准确地分析淋巴管密度。整片组织的免疫染色很理想，毛细淋巴管盲端、毛细淋巴管网和淋巴管丛清晰可见。 结论 整片组织免疫染色能够很好地显示组织内完整的淋巴管构筑。
Objective To reveal lymphatic vessels in the tissue with the whole-mount immunostaining and compare this method with indirect injection and section immunostaining methods. Methods The lymphatic vessels in the capsule of the newborn liver were indirectly injected with Prussian blue; the architecture of the superficial hepatic lymphatic vessels was viewed. Drainage of the vessels was analyzed with collodion sections. Human skin of the femoral region was removed, and the distribution of the lymphatic vessels in paraffin and frozen sections was determined. After the skin of the ear and back, diaphragm and small intestine were removed from rats, the tissue pieces were stained with immunofluorescence, and the architecture of the lymphatic vessels in the tissue was examined.
Results Indirect injection of Prussian blue revealed the architectures and drainage of the lymphatic vessels, while staining of the lymphatic vessels was short of specificity. With immunostaining, the tissue sections showed the distribution of the lymphatic vessels. However, the density of the lymphatic vessels was not analyzed accurately in this method. Whole-mount immunostaining of the tissues was desirable. The blind ends of the lymphatic capillaries, lymphatic capillary network and lymphatic plexus were clear. Conclusion The architecture of the intact lymphatic vessels in the tissue may be demonstrated well with the whole-mount immunostaining.
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国家自然科学基金项目;教育部高等学校博士学科点专项基金;上海市科委国际合作基金
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