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减少本地冷生物样本中的SEM充电工件

Abner Velazco¹, Thomas Glen¹, Sven Klumpe²

¹The Rosalind Franklin Institute, Harwell Science & Innovation Campus, Didcot, UK.

Nature communications

|June 4, 2025

概括

在冷水合生物样本的扫描电子显微镜 (SEM) 中的充电器件阻碍了成像. 我们引入交联扫描来克服这些局限性,揭示以前隐藏的细胞结构.

科学领域:

细胞生物学细胞生物学
显微镜技术显微镜技术
生物物理学的生物物理.

背景情况:

在冷水合样本上的扫描电子显微镜 (SEM) 提供了与原生细胞结构相似的中等尺度成像.
串联聚焦离子束/扫描电子显微镜 (FIB/SEM) 能够对细胞和组织进行3D重建.
非导电生物样本的SEM中的充电工件限制了成像分辨率和细节.

研究的目的:

开发和演示一种新的扫描方法,以减轻冷水化生物样本在SEM中的充电器件.
为了使以前模糊的亚细胞结构和细胞区块可视化.
将FIB/SEM技术的适用性扩展到更广泛的非导电生物标本.

主要方法:

在串行FIB/SEM采集过程中实施交叉扫描策略.
该方法应用于冷保存的生物样本,包括哺乳动物细胞,藻类和老鼠大脑组织.
通过常规扫描与交叶扫描获得的成像结果的比较分析.

主要成果:

交叉扫描有效地减弱了充电器件,显著提高了图像质量和功能可见性.
之前无法观察到的结构,如膜接触点,藻类降解区和轴突膜网络,已成功可视化.
该方法允许更清晰的3D重建,并促进了与高分辨率冷电子显微镜 (cryoEM) 分析的相关性.

结论:

交叶扫描是一种强大的技术,用于克服冷水化生物样本的SEM中的充电器件.
这种方法增强了FIB/SEM的实用性,用于对非导电生物标本的详细结构分析.
这种方法对于在各种生物研究领域推进细胞和组织成像具有广泛的意义.

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