3D相关冷结构照明光和软X射线显微镜阐明了reovirus细胞内释放途径
Ilias Kounatidis1, Megan L Stanifer2, Michael A Phillips3
1Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 0DE, UK.
Cell
|July 2, 2020
概括
研究人员开发了一个3D相关成像平台, 结合了超分辨率显微镜和软X射线断层对冷保存的细胞. 这使得细胞结构和过程的高分辨率可视化,如病毒释放,弥合宏观和微观尺度.
科学领域:
- 生物物理
- 细胞生物学
- 显微镜
背景情况:
- 相关成像需要对齐宏观和微观特征.
- 冷保存对于X射线成像至关重要,但通常会限制光显微镜的分辨率.
- 现有的方法难以与X射线和可见光成像相匹配.
研究的目的:
- 开发一个用于冷条件下的细胞3D相关成像的综合平台.
- 为了弥合X射线成像和超高分辨率光显微镜之间的分辨率差距.
- 能够在现场对细胞过程进行详细的超结构分析.
主要方法:
- 开发一个集成的,用户友好的3D相关成像平台.
- 使用超分辨率结构化照明显微镜 (SR-SIM).
- 将SR-SIM与软X射线断层扫描 (SXT) 结合在玻璃体冰中的冷保存细胞.
主要成果:
- 展示了在玻璃体冰中的高分辨率3D成像平台.
- 成功将超分辨率光显微镜数据与SXT数据相关联.
- 从细胞内囊泡释放的早期阶段的可视化.
结论:
- 开发的平台克服了相关冷成像的分辨率限制.
- 能够在超结构层面详细研究病毒感染动态和病毒诱导的结构.
- 为需要多尺度成像的细胞生物学研究提供了强大的工具.
相关概念视频
Cryo-electron Microscopy
4.0K
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...
4.0K
Three-Dimensional Microscopy in Microbiology
675
Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...
675
Immunofluorescence Microscopy
12.6K
A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
12.6K


