STED microscope

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We apply for financial support to purchase a STED microscope. STED microscopy is an optical super-resolution method that is capable of generating fluorescence images of biological samples with a spatial resolution of a few tens of nanometers. STED is a true-optical super-resolution method without elements of computational image processing. STED is capable of 3D imaging of thick samples through the integration of e.g. adaptive optics and allows for live-cell imaging. Our research group has recently contributed to STED microscopy by implementing the concept of “exchangeable fluorophore labels”, which we adapted from single-molecule localization microscopy. Such labels transiently and repeatedly bind to a target, which is achieved by keeping these labels available in the imager buffer. At the first place, a continuous exchange of labels to and from their target minimizes photobleaching in any fluorescence imaging technique. Applied to STED microscopy, additional benefits are: easy-to-implement multi-color imaging, 3D volume imaging and live cell imaging, which all profit from reduced photobleaching. Here, we apply for a STED microscope to further develop the technology into a structural cell biology microscopy method by enabling multi-protein imaging by using novel concepts of protein labeling. This work will focus on single cells, multi-cellular organisations and tissue sections. Second, we will develop quantitative STED microscopy, again profiting from and adapting our knowledge in single-molecule localization microscopy and integrating new protein labels. Third, we will develop computational pipelines that allow integrating various aspects of state-of-the-art deep-learning assisted microscopy tools, e.g. denoising for low light dose imaging of live specimens for a prolonged observation time. These work packages demand for a high-performance STED microscope equipped with multiple excitation sources, a modular construction and the option of integrating novel optical or auxiliary elements to the microscope system.