STED microscope

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.

DFG Programme Major Research Instrumentation

Major Instrumentation STED Mikroskop

Instrumentation Group 5090 Spezialmikroskope

Applicant Institution Goethe-Universität Frankfurt am Main

Leader Professor Dr. Mike Heilemann