Correlative cryogenic fluorescence and soft x-ray tomography of cells

该项目的主要目标是将软X射线断层扫描和荧光显微镜(两者都在低温环境中)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，细胞成像不会因为使用造影剂、脱水、固定和切片而产生潜在的伪影。将特别强调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，那里是世界领先的生命科学研究软X射线显微镜的所在地。由于荧光显微镜与X射线显微镜相比，空间分辨率较低(100 nm比20 nm)，所以我使用了结构照明荧光显微镜。这项技术与软x射线断层成像技术的结合，将为一侧的标记分子和另一侧天然水合细胞的亚细胞结构提供前所未有的高分辨率3D成像。

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.