| 高压快速冷冻技术在两种培养细胞超微结构中的应用 |
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| 引用本文： | 肖丽国,李茵茵,李红梅,张雪晴,贾旭东,张勤奋.高压快速冷冻技术在两种培养细胞超微结构中的应用[J].电子显微学报,2017,36(2). |
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| 作者姓名： | 肖丽国 李茵茵 李红梅 张雪晴 贾旭东 张勤奋 |
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| 作者单位： | 中山大学生命科学学院,有害生物控制与资源利用国家重点实验室,广东 广州510275 |
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| 基金项目： | 国家自然科学基金资助项目，NSFC-RGC资助项目 |
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| 摘 要： | 高压快速冷冻(high-pressure freezing,HPF)能使样品在高压下毫秒内从室温降到液氮温度,所有分子瞬间固定在原位,随后启用冷冻置换(freeze substitution,FS),在低温下缓慢固定的同时用有机溶剂替代样品中的水分,从而保存样品的细微结构接近自然状态.本文使用高压快速冷冻/冷冻置换方法对草地贪夜蛾细胞(sf9细胞)和感染沙眼衣原体的小鼠成纤维细胞(McCoy细胞)进行电镜制样观察,同时对比传统化学固定两种细胞超微结构的差异.结果发现,利用HPF/FS技术,两种细胞中的膜结构、细胞骨架结构更能良好地保存,核膜、高尔基体和囊泡膜平滑完整,无皱缩或断裂,细胞形态更接近自然状态.
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| 关 键 词： | 高压快速冷冻 冷冻置换 化学固定 McCoy细胞 sf9细胞 |
The application of high-pressure freezing in the ultrastructures of two cultured cells |
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| XIAO Li-guo,LI Yin-yin,LI Hong-mei,ZHANG Xue-qing,JIA Xu-dong,ZHANG Qin-fen.The application of high-pressure freezing in the ultrastructures of two cultured cells[J].Journal of Chinese Electron Microscopy Society,2017,36(2). |
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| Authors: | XIAO Li-guo LI Yin-yin LI Hong-mei ZHANG Xue-qing JIA Xu-dong ZHANG Qin-fen |
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| Abstract: | High-pressure freezing ( HPF ) can instantly fix all molecules in situ by decreasing the temperature from ambient temperature to about -196 ℃ in milliseconds. The following freeze substitution ( FS) can proceed with the dehydration at a low temperature. In this paper, the applications of HPF/FS in fixing the sf9 cells and the McCoy cells infected with Chlamydia trachomatis were reported. The results reveal that the cytoplasm, membrane structures, Golgi apparatus, vesicles and micro fiber structures in the cells with HPF/FS fixation are preserved better than those in the cells with chemical fixation. |
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| Keywords: | high-pressure freezing freeze substitution chemical fixation McCoy cell sf9 cell |
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