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。
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, immunocytochemistry, immunofluorescent labeling and quantum dot labeling under light microscope and the locations were marked on the chamber slides. Whereafter, the chamber slides 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 infected cells under light microscope can be successfully relocated and observed on the ultrathin section under transmission electron microscope.Conclusions: A simple and feasible CLEM for virus detection was established based on Lab-Tek chamber slide.
作者
冯红丽
彭宇宁
宋敬东
FENG Hong-li;PENG Yu-ning;SONG Jing-dong(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;School of Physics and Electronics,Hunan Normal University,Changsha Hunan 410082,China)
出处
《电子显微学报》
CAS
CSCD
北大核心
2023年第1期86-93,共8页
Journal of Chinese Electron Microscopy Society
基金
国家生物安全实物核心资源库支撑关键技术研究(No.2022YFC2602202)。
关键词
透射电子显微镜
光镜-电镜联用技术
病毒
检测
transmission electron microscope
correlative light and electron microscopy
virus
detection
