一种用于贴壁培养细胞感染病毒检测的简易光镜-电镜联用技术

冯红丽,彭宇宁,宋敬东*

一种用于贴壁培养细胞感染病毒检测的简易光镜-电镜联用技术

冯红丽,彭宇宁,宋敬东*

(1.中国疾病预防控制中心病毒病预防控制所,传染病预防控制国家重点实验室,北京 100052;2. 湖南师范大学物理与电子科学学院,湖南 长沙 410082)

摘  要  目的:建立一种简单易行的适用于病毒检测的光镜-电镜联用技术。方法:使用Lab-Tek腔室玻片培养细胞并接种不同类型病毒,通过光学显微镜选取目标病变细胞或通过荧光蛋白、免疫细胞化学标记、免疫荧光标记、量子点标记确定目标细胞并在腔室玻片上标记其部位。随后将腔室玻片改造为包埋模具,最后钻取目标细胞及其包埋树脂,并对目标细胞进行靶向性超薄切片及透射电子显微镜观察。结果:光镜下选取的目标细胞在透射电子显微镜下可顺利定位并进行观察。结论:建立了一种基于腔室玻片包埋模具的可用于病毒检测的光镜-电镜联用技术。

关键词  透射电子显微镜;光镜-电镜联用技术;病毒;检测

中国分类号:R-331;R372;Q934.2 文献标识码:A      doi:10.3969/j.issn.1000-6281.2023.01.013

 

A simple correlative light and electron microscopy for virus detection in adherent culture cells

FENG Hong-li1, PENG Yu-ning12, SONG Jing-dong*1

(1. State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052;

2. School of Physics and Electronics, Hunan Normal University, Changsha Hu’nan 410082, China)

Abstract     Objective: To establish a simple correlative light and electron microscopy (CLEM) for virus detection. Methods: Cells were cultured in Lab-Tek chamber slides and different types of viruses were inoculated in the cells respectively. Target cells infected were identified by cytopathic effect (CPE), fluorescent protein, immunohistochemistry, immunofluorescent labeling and quantum dot labeling under light microscope and the locations were labeled on the chamber slides. Whereafter, the chamber slides which adhered cells were transformed into embedding moulds. Finally, the target cells and the embedding resin were drilled out vertically with a designed hole cutter and ultrathin sectioned. Counterpart targeted cells were observed under electron microscope.Results: The target cells infected under light microscope can be successfully relocated and observed on the ultrathin section under transmission electron microscope.Conclusion: A simple and feasible CLEM for virus detection was established based on Lab-Tek chamber slide.

Kaywords      transmission electron microscope;correlative light and electron microscopy;virus;detection

 

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