Superresolution stimulated emission-depletion (STED) microscope

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Superresolution fluorescence microscopy is needed in order to investigate cellular structures and dynamic cellular processes below the Abbe diffraction limit.Accordingly, the requested stimulated emission-depletion (STED) microscope will serve the need for superresolution light microscopy on a nanometer-scale (nanoscopy) of several research groups at the Institute for Biology, Humboldt-Universität zu Berlin. No similar instrumentation using the STED technique is available at the Humboldt-Universität zu Berlin.At the moment, superresolution microscopy experiments at the Institute for Biology, Humboldt- Universität zu Berlin are only possible in collaboration with external institutions, or using a photoactivated localization microscope (PALM) of the research group ‘Molecular Biophysics’ of Prof. A. Herrmann. However, special photo-activatable fluorophores and specific experimental conditions are needed for PALM, which frequently requires re-construction of available fluorescent protein fusion constructs. In addition, the available PALM microscope is used to capacity by the research group ‘Molecular Biophysics’ of Prof. Herrmann and the newly established working group ‘Experimental Biophysics / Mechanobiology’ of junior Prof. E. Klotzsch. Importantly, no other live cell-capable superresolution microscope is available at the Humboldt-Universität zu Berlin and thus many cutting-edge research questions in microbial biology, infection biology, cell biology, neurobiology and biophysics can currently only be addressed in external collaborations. This is not optimal to achieve continuity, sustainability and efficiency in any ongoing and future research in the life sciences at the Humboldt-Universität zu Berlin. The attached research projects of the main applicants Prof. M. Erhardt and Prof. C. Schmitz-Linneweber, the co-applicants Dr. B. Beckmann (independent junior research group), Prof. M. Brecht, Prof. P. Hegemann, Prof. A. Herrmann/Dr. T. Korte, Prof. E. Klotzsch, Dr. S. Reber (independent junior research group) and Dr. N. Tschowri (Emmy Noether junior research group) highlight the wide variety of research questions and applications that require imaging techniques beyond the diffraction limit. Accordingly, the requested STED microscope will serve the need for superresolution imaging in many ongoing and planned research projects of several research groups at the Institute of Biology. The ability to perform STED microscopy will propel ongoing projects to a new level of spatial understanding of biological structures and will also allow the development of entirely new research lines. The wealth of projects, both running and proposed, will ensure operation of the requested microscope to full capacity. The procurement of a STED microscope will thus be instrumental for continued competitive research at the Institute of Biology and, not the least, will enable a large number of local, national and international collaborations.