Correlative cryogenic fluorescence and soft x-ray tomography of cells

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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.