全组织包埋免疫荧光染色技术在小鼠淋巴管形态研究中的应用  

The application of Whole-Mount immuno fluorescence staining technique in the study of lymphatic morphology in mice

作　　者：孙笛[1] 袁兆琪 赵蒙蒙 倪涛[1] 于子尤 刘宁飞[1] 杨军[1] Sun Di;Yuan Zhaoqi;Zhao Mengmeng;Ni Tao;Yu Ziyou;Liu Ningfei;Yang Jun(Department of Plastic and Reconstructive Surgery,Shanghai Ninth People′s Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200001,China;Radiology Department,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,China)

机构地区：[1]上海交通大学医学院附属第九人民医院整形外科,200001 [2]郑州大学附属郑州中心医院放射科,450007

出　　处：《中华整形外科杂志》2019年第7期681-685,共5页Chinese Journal of Plastic Surgery

基　　金：国家自然科学基金(81272146).

摘　　要：目的探讨全组织包埋免疫荧光染色技术在淋巴管荧光染色的特点和优势.方法从正常C57小鼠上分别获取耳部皮肤组织、后肢淋巴管和肠系膜淋巴管,将这些组织样本进行全组织包埋免疫荧光染色,经多聚甲醛固定、封闭液封闭、一抗二抗不同时间孵育,在共聚焦激光显微镜下观察小鼠毛细淋巴管和集合淋巴管的解剖结构.结果经过全组织包埋免疫荧光染色的耳部皮肤能清晰显示毛细淋巴管内皮细胞及其盲端,后肢集合淋巴管和肠系膜淋巴管还能清晰地显示淋巴管及其平滑肌和瓣膜.结论全组织包埋免疫荧光染色技术既能清晰、立体地观察到淋巴管外部形态,又能深入细微地观察淋巴管内部结构,从而更精准地研究淋巴管的生理病理改变.Objective To elaborate the characteristics and advantages of Whole-Mount immune fluorescence staining by observing the lymphatic vessels of mice. Methods The ear skin tissue, the hindlimb lymphatic vessels and the mesenteric lymphatic vessels were harvested from normal C57 mice. The tissue samples were subjected to whole-tissue immunofluorescence staining.These tissue samples were fixed by paraformaldehyde, blocked by bovine serum and incubated in primary and secondary antibodies. Then, the lymphatic vessels were observed and analyzed in these samples with a confocal laser-scanning microscope. Results The capillary lymphatic vessels and lymphatic endothelial cells can be clearly showed in the ear skin. The valves and smooth muscles can be clearly showed in the hindlimb and mesenteric lymphatic vessels by Whole-Mount immunofluorescence staining. Conclusions The whole-tissue immunofluorescence staining technique can observe the external morphology of lymphatic vessels clearly and stereoscopically, and can deeply observe the internal structure of lymphatic vessels. This technique can provide more accurate study on physiology and pathology of lymphatic vessels.

关 键 词：免疫荧光染色 淋巴管 淋巴管内皮细胞 

分 类 号：R361.2[医药卫生—病理学]