超薄冷冻切片在感染的非洲绿猴肾细胞培养物中定位流感病毒抗原的应用。

The use of ultrathin cryosections for localisation of influenza virus antigens in infected vero cell cultures.

作者信息

Beesley J E, Campbell D A

出版信息

Histochemistry. 1984;80(5):497-501. doi: 10.1007/BF00495441.

Abstract

The localisation of influenza virus antigens in infected Vero cell monolayer cultures by post embedding immunoelectron microscopy requires both good resolution and the retention of antigenicity in the tissue sections. Ultrathin cryosections are superior to ultrathin resin sections for this purpose. The colloidal gold probe was used in conjunction with specific antibody preparations to localise three viral proteins. Antibody raised against haemagglutinin glycoproteins labelled the host cell membrane and the virus fringe without contamination of the host cell nucleus, whereas antibody raised against viral nuclear protein labelled throughout the host cell cytoplasm and nucleus. Matrix protein was localised within the nucleus and was associated with the host cell membrane of the infected cell. The appearance of all these proteins was maximal 24 h post infection.

摘要

通过包埋后免疫电子显微镜对感染的非洲绿猴肾细胞单层培养物中的流感病毒抗原进行定位，既需要良好的分辨率，又需要在组织切片中保留抗原性。为此，超薄冷冻切片优于超薄树脂切片。胶体金探针与特异性抗体制剂联合使用，以定位三种病毒蛋白。针对血凝素糖蛋白产生的抗体标记了宿主细胞膜和病毒边缘，而未污染宿主细胞核，而针对病毒核蛋白产生的抗体则在整个宿主细胞质和细胞核中进行标记。基质蛋白定位于细胞核内，并与感染细胞的宿主细胞膜相关。所有这些蛋白在感染后24小时出现的量最大。