相关实验视频

Updated: Sep 14, 2025

Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy

Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy

Published on: November 6, 2021

基本温度,浸泡介质和EM电网材料对低温光学超分辨率显微镜中的脱门的影响

Soheil Mojiri¹, Joseph M Dobbs², Niko Faul³

¹Molecular Systems Biology (MSB) unit, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, Heidelberg, 69117, Germany.

Journal of structural biology

|July 19, 2025

概括

优化低温单分子局部化显微镜 (cryo-SMLM) 需要了解样本脱. 金属支和冷浸泡介质增强了散热,使得更高的激光强度可以改善冷CLEM成像.

科学领域:

结构生物学是结构生物学.
显微镜技术的使用方法
生物物理学的生物物理.

背景情况:

低温相对光和电子显微镜 (cryo-CLEM) 将分子标记与高分辨率结构数据相结合.
在光显微镜和电子显微镜 (EM) 之间存在显著的分辨率差距.
低温单分子定位显微镜 (cryo-SMLM) 旨在弥合这一差距,但面临着激光诱导样品脱的挑战.

研究的目的:

调查在冷SMLM过程中影响样本脱维的因素.
为了确定最大限度地减少除化和最大限度地提高激光耐受性的最佳条件.
为提高冷SMLM设置性能提供定量见解.

主要方法:

有限元模拟用于模拟热传递和脱玻璃化.
模拟预测的实验验证.
系统评估基冷却温度,浸泡介质和EM电网支材料 (碳与金属).

主要成果:

低温浸泡介质可以改善碳支物的散热.
金属支 (金,银涂) 可以承受更高的激光强度,特别是在冷的气或真空中.
金色支显示在640nm照明下高激光值.
金属支在真空冷静态中表现出高效的散热效率.

结论:

基本冷却温度,浸泡介质和支材料极大地影响了冷SMLM激光的耐受性.
金属支和冷浸泡介质是缓解脱的关键.
使用这些发现的优化冷SMLM设置可以改善冷CLEM成像分辨率和数据质量.

关键词:

基本温度基本温度基本温度克里奥-克莱姆 (Cryo-CLEMEM) 是一种低温电子断层扫描 (Cryogenic electron tomography) 是一种低温的电子断层扫描技术.达维特的分化方式光显微镜的光学显微镜.单分子局部化显微镜.

更多相关视频

Preparation of High-Temperature Sample Grids for Cryo-EM

Preparation of High-Temperature Sample Grids for Cryo-EM

Published on: July 26, 2021

Manual Blot-and-Plunge Freezing of Biological Specimens for Single-Particle Cryogenic Electron Microscopy

Manual Blot-and-Plunge Freezing of Biological Specimens for Single-Particle Cryogenic Electron Microscopy

Published on: February 7, 2022

相关概念视频

Cryo-electron Microscopy

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...