高压冷冻及冷冻替代技术在不同生物样品超微结构中的应用  

The application of highpressure freezing-freeze substitution technique in ultrastructure of different biological samples

作　　者：葛灵 钦风凌 祁方杰 涂溢晖 何钧 边玮 GE Ling;QIN Feng-ling;QI Fang-jie;TU Yi-hui;HE Jun;BIAN Wei(Center for Excellence in Molecular Cell Science,State Key Laboratory of Cell Biology,Chinese Academy of Sciences,Shanghai 200031,China)

机构地区：[1]中国科学院分子细胞科学卓越创新中心,细胞生物学国家重点实验室,细胞分析技术平台电镜技术部门,上海200031

出　　处：《电子显微学报》2021年第1期61-66,共6页Journal of Chinese Electron Microscopy Society

基　　金：中国科学院仪器设备功能开发项目.

摘　　要：高压冷冻仪在固定样品时,高压和冷冻基本是同步发生的,使得生物样品在数毫秒的时间内被瞬时固定,保证其内部结构接近于生理状态,有效减少了化学固定法中的化学试剂对细胞结构的破坏,极大程度保存了细胞在近似自然状态下的超微结构。本文分别采用化学固定法(CF)和高压冷冻固定法(HPF)对小鼠的睾丸组织、Hela细胞和酵母进行了样品制备,旨在比较这两种方法对不同生物样品超微结构的影响,促进高压冷冻-冷冻替代(HPF-FS)技术在生物学中的应用。透射电镜(TEM)观察结果显示:与传统的化学固定相比,高压冷冻固定可以清晰、完整地保存生物样品的膜结构和亚细胞器结构,细胞形态更加接近自然。During the process of fixing the sample, the high pressure and freezing basically occur simultaneously. The biological sample is instantaneously fixed within a few milliseconds to ensure that its internal structure is close to the physiological state, which effectively reduces the damage to the cell structure caused by the chemical reagent in the chemical fixation method and greatly preserves the ultrastructure of the cell in the approximate natural state. In this paper, we compared chemical fixation method(CF) and high pressure freezing(HPF) method in preparation samples of mouse testicular tissue, Hela cells and yeast to figure out the influence in different biological samples ultrastructure and to promote the application of high pressure freezing-freeze substitution(HPF-FS) technique in biology. Transmission electron microscopy(TEM) observation results demonstrated that compared with traditional chemical fixation, high pressure freezing fixation could clearly and completely preserved the membrane structure and suborganelle structure of biological samples, and the cell morphology was closer to nature.

关 键 词：高压冷冻-冷冻替代 超微结构 睾丸 HELA细胞 酵母 

分 类 号：Q-336[生物学] Q249