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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
引用本文:葛灵,钦风凌,祁方杰,涂溢晖,何钧,边玮.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[J].电子显微学报,2021,40(1):61-66.
作者姓名:葛灵  钦风凌  祁方杰  涂溢晖  何钧  边玮
作者单位: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;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;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;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;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;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
基金项目: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...

关 键 词:高压冷冻-冷冻替代  超微结构  睾丸  Hela细胞  酵母

The application of highpressure freezing-freeze substitution technique in ultrastructure of different biological samples
GE Ling,QIN Feng-ling,QI Fang-jie,TU Yi-hui,HE Jun,BIAN Wei.The application of highpressure freezing-freeze substitution technique in ultrastructure of different biological samples[J].Journal of Chinese Electron Microscopy Society,2021,40(1):61-66.
Authors:GE Ling  QIN Feng-ling  QI Fang-jie  TU Yi-hui  HE Jun  BIAN Wei
Institution:(Center for Excellence in Molecular Cell Science,State Key Laboratory of Cell Biology,Chinese Academy of Sciences,Shanghai 200031,China)
Abstract: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.
Keywords:high pressure freezing-freezing substitution  ultrastructure  testis  Hela cell  yeast
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