Transformative microscopes to image across spatiotemporal scales

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Project abstract How molecular organization and activity leads to tissue level outcomes is, arguably, one of the big remaining open questions in biology. Bridging these two areas is key to biomedical advances, yet is technically challenging because we cannot observe with molecular precision at the tissue scale. While optical microscopy is the method of choice to observe architecture and dynamics within living cells and organisms, it has fundamental limitations in spatiotemporal resolution and optical penetration depth. Thus our most detailed observations of cellular dynamics and ultrastructure have been limited to single cells that were far removed from their physiological context. Here I propose to significantly expand the reach of super-resolution microscopy to encompass tissues and whole model organisms. This lies far outside of the capabilities of the current state of the art and requires significant progress in volumetric acquisition speed, sensitivity and optical penetration depth. I propose to advance the field by combining three different fields of microscopy. Some of these combinations are non-trivial and thus have not been experimentally tractable so far. I propose novel concepts that can bridge these different fields and overcome technical and fundamental limitations. Furthermore, a novel adaptive sampling approach that only images the most informative voxels at the highest resolution will overcome fundamental barriers to high-resolution imaging over large volumes. I hypothesize that my new instrumentation combined with an intelligent sampling strategt can improve acquisition speed up to 100 fold while reducing phototoxicity and data storage needs. The resulting new microscope technology will dramatically speed up high-resolution imaging over large volumes and hence will enable large volume imaging experiments that have been prohibited by either lack of spatial resolution, optical penetration depth or acquisition speed.