Correlative cryogenic fluorescence and soft x-ray tomography of cells

细胞的相关低温荧光和软 X 射线断层扫描

• 批准号: 355191992
• 负责人: Dr. Venera Weinhardt
• 金额: --
• 依托单位: Centre for Organismal Studies (COS)
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/355191992?language=en
• 关键词: Correlative; cryogenic; fluorescence; soft; ray

The primary goal of the project is to correlate soft x-ray tomography and fluorescence microscopy (both in cryogenic environment) in order to link molecular events and ultrastructure of single eukaryotic cells. With such an approach, cells are imaged free from potential artifacts due to use of contrast enhancing agents, dehydration, fixation, and sectioning. Special emphasis will be put on obtaining similar spatial resolution in both methods. I plan to realize this project at the Lawrence Berkeley National Laboratory, where the worlds leading soft x-ray microscope for life science research is located. Since fluorescence microscopy, in comparison to x-ray microscopy, suffers from poor spatial resolution (100nm versus 20nm), I employ structured illumination fluorescence microscopy. The combination of this technique with soft x-ray tomography will provide unprecedented high resolution 3D imaging of labeled molecules on the one side and of subcellular structures of natively hydrated cells on the other side.

该项目的主要目标是将软x射线断层扫描和荧光显微镜（均在低温环境下）联系起来，以便将单个真核细胞的分子事件和超微结构联系起来。通过这种方法，细胞成像没有由于使用造影剂、脱水、固定和切片而产生的潜在伪影。特别强调的是在这两种方法中获得相似的空间分辨率。我计划在劳伦斯伯克利国家实验室实现这个项目，那里有世界领先的生命科学研究软x射线显微镜。由于与x射线显微镜相比，荧光显微镜的空间分辨率较差（100nm与20nm），因此我采用了结构照明荧光显微镜。该技术与软x射线断层扫描相结合，将提供前所未有的高分辨率3D成像，一边是标记分子，另一边是天然水合细胞的亚细胞结构。