使用cRNA探针的原位杂交方法与免疫组织化学双重标记技术在皮肤石蜡切片中的应用探讨  

Application of double-lableling technique between in situ hybridization with cRNA probe and immunohistochemistry in the paraffin section of skin

作　　者：杨怡[1] 魏燕燕[2] 高剑[1] 武胜昔[2] 高天文[1] 

机构地区：[1]第四军医大学西京医院全军皮肤病研究所,陕西西安710032 [2]第四军医大学解剖学教研室暨梁銶琚脑研究中心,陕西西安710032

出　　处：《中国美容医学》2009年第4期503-506,共4页Chinese Journal of Aesthetic Medicine

摘　　要：目的:探讨使用cRNA探针的原位杂交技术结合免疫组织化学双重标记的方法在皮肤石蜡切片中应用的可行性。方法:在20例痤疮及毛囊炎患者石蜡切片上,根据痤疮丙酸杆菌16sRNA序列设计合成cRNA探针,优化探针穿透条件,进行原位杂交,并使用鼠抗人CD1a单克隆抗体同时对表皮朗格汉斯细胞进行免疫组化标记。显微镜观察双标信号的共存状态。结果:在20例痤疮患者皮肤石蜡切片的毛囊上皮,皮脂腺及导管部位获得蓝紫色原位杂交阳性信号,同时在表皮及皮脂腺处可见明确的棕黄色树突状阳性信号。结论:使用地高辛标记的cRNA探针进行原位杂交,结合免疫组织化学双重标记的方法能够在皮肤石蜡切片上对微生物及表皮细胞成分进行准确定位。Objective To explore the double-labeling method of in situ hybridization （ISH） with cRNA probe and immunohistochemistry in the paraffin sections of the skin. Methods According to the sequence of Propionibacterium acnes 16sRNA, the cRNA probe was designed and synthesized, after optimizing the conditions for probes penetrating, in situ hybridization was carried out on paraffin sections of patients with acne, then, the mouse anti-human monoolonal antibody CDla was used for Langerhans cell immunohistochemistry. Finally, the co-existence of the two signals was observed under the microscope. Results The blue-violet positive signals of ISH can be observed in the hair follicle epithelium, sebaceous glands and ducts of paraffin section of skin. Meanwhile, the brown dendritic positive signal also can be seen in the sebaceous glands. Conclusions The use of in situ hybridization with digoxin-labeled cRNA probe combined with immunohistochemistry can locate microbial component and epidermal cells on skin paraffin section exactly.

关 键 词：原位杂交 免疫组织化学 痤疮丙酸杆菌 朗格汉斯细胞 

分 类 号：Q813.1[生物学—生物工程]