尽量减少冰污染在试样准备过程中的冷软X射线断层扫描和冷电子断层扫描
Chia-Chun Hsieh1, Zi-Jing Lin1, Lee-Jene Lai1
1Experimental Division, Synchrotron Radiation Research Center, 101 Hsin-Ann Road, Hsinchu Science Park, Hsinchu 300092, Taiwan, ROC.
Journal of structural biology: X
|November 5, 2024
概括
一个新的冷多网格盒存储系统在冷软X射线断层扫描 (cryo-SXT) 的样本准备过程中最大限度地减少了冰污染. 这种高效的方法保留了样品完整性,用于高分辨率的3D细胞成像.
科学领域:
- 细胞和分子成像技术
- 生物物理学的生物物理.
- 冷电子显微镜和断层扫描
背景情况:
- 冷软X射线断层扫描 (cryo-SXT) 能够在近原生状态下对细胞进行3D成像.
- 高效的冷样本网准备对于最大限度地减少冰污染和最大限度地提高光束时间效用至关重要.
- 目前的方法可能容易形成冰块,可能会损害样品质量.
研究的目的:
- 为冷样本网设计和评估一个改进的储存系统.
- 为了减少冷SXT和冷电子断层扫描 (cryo-ET) 样品的准备过程中的冰污染.
- 为了提高冷样本网准备的效率和可靠性.
主要方法:
- 开发一种新型的冷式多网格盒存储系统,包括一个外,漏斗支架和多网格盒容器.
- 该系统应用于A549细胞冷网准备.
- 使用冷光显微镜和冷SXT进行相关成像,以评估冰的形成和样品质量.
主要成果:
- 化多网格盒存储系统成功增加了冷样本网的临时存储容量.
- 该系统减少了网箱转移的频率,大大降低了冰污染的可能性.
- 相对成像证实了新系统准备的网格上有限的冰形成,使线粒体晶体的清晰3D可视化成为可能.
结论:
- 开发的冷多网格盒存储系统有效地减少了冷SXT样品准备过程中的冰污染.
- 该系统提高了为高分辨率成像准备冷样本网的效率和可靠性.
- 该系统适用于冷SXT和冷电子断层扫描 (冷ET) 工作流程.
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