三种显微技术对人毛囊蠕形螨的观察和研究  

Observation and Study of Human Demodex folliculorum Using Three Kinds of Micro-Detection Technology

作　　者：佘俊萍[1] 张锡林[2] 王光西[1] 荣华[1] 牛慧[1] 王利芳[1] 

机构地区：[1]泸州医学院病原生物学教研室,四川泸州646000 [2]第三军医大学病原生物学教研室,重庆400038

出　　处：《四川动物》2011年第1期47-49,F0004,共4页Sichuan Journal of Zoology

基　　金：泸州医学院2010年课题(川南地区人蠕形螨基因组DNA的提取和检测以及虫体抗原对上皮细胞炎症损伤机制的研究;编号767);2011年四川省卫生厅课题(川南地区人蠕形螨基因组DNA遗传多态性和致病性研究;编号100245)

摘　　要：目的采用荧光显微镜、扫描电镜和激光共聚焦显微镜对人体毛囊形蠕形螨进行形态学观察。方法选用5μg/mL碘化丙啶(propidine iodide,PI)对虫体进行荧光染色,避光染色15min,分别置于荧光显微镜和激光共聚焦显微镜下观察;2.5%戊二醛固定虫体标本,梯度酒精和叔丁醇脱水,金喷镀后扫描电镜观察。结果荧光显微镜下碘化丙啶对虫体有很强的结合力,虫体荧光信号均匀展示于细胞表面,充分展现虫体形态,扫描电镜更加清楚、细致地展示人体毛囊型蠕形螨的超微结构,激光共聚焦显微镜将虫体分层扫描图片进行三维重建,真实、完全、直观地展露了虫体。结论三种显微技术均可展示蠕形螨超微形态。PI对虫体有很好的荧光染色作用,使激光扫描共聚焦显微镜能够获得更加精准的超微形态结构,结合三维重建技术有着广泛的应用价值。Objective To observe the morphological characteristics of Demodex folliculorum by using fluorescence microscopy, SEM and confocal laser scanning microscopy. Methods To observe and photograph Dernodex folliculorum by using fluorescence microscopy and confocal laser scanning microscopy after fluorescent staining with propidine iodide for 15 minutes. To observe and photograph Demodexfolliculorum by using SEM after fixation with 2.5% glutaral, dehydration of gradient ethanol and tert-butyl alcohol and spray plating. Results The specimens combined with propidine iodide well. Fluorescence signals were distributed uniformly on the surfaces of the specimens. The specimens could be observed clearly with a fluorescence microscope. The ultrastructure of Demodex folliculorurn was displayed using SEM. High definition three-di- mensional pictures were obtained by using confocal laser scanning microscopy and finished three-dimensional reconstruction, Demodex folliculorum can be true, complete and intuitive exposed. Conclusion Three kinds of micro technologies can be used to show Demodex folliculorum. PI has a good fluorescent dyeing effect on Demodex folliculorum. CLSM can obtain a more precise uhrastructure and combined with three-dimensional reconstruction technology, it is quite a valuable method.

关 键 词：毛囊形蠕形螨 碘化丙啶 荧光显微镜 激光共聚焦显微镜 扫描电镜 

分 类 号：Q954[生物学—动物学]