Nanoscopic investigation of dynamic subcellular events in health and disease using TauSTED technology

Many fundamental biological processes are highly dynamic and occur at the nanometer scale. For example, tiny units like proteins and molecules (<30nm in diameter) continuously move between different subcellular compartments (30-100nm in diameter). Neuronal synapses (~20-40nm in diameter), which are key sites of brain communication, are constantly reshaped by very thin, mobile extensions of neighbouring cells. The ability to directly investigate these events in real-time and to visualise how they are disrupted during pathologies have tremendous potential to advance biomedical research into human diseases. Several technological advancements have pushed boundaries beyond the diffraction limit of light (~200nm), including Stimulated Emission Depletion (STED) super-resolution microscopy. However, its application is limited, as many delicate live biological samples and labelling methods are incompatible with the required high laser power. The Leica TauSTED Xted (TauSTED) microscope is the latest revolutionising technology, combining STED with novel fluorescence lifetime-based detection mechanisms to overcome this challenge by effectively removing the requirement of high laser power. The system enables researchers to finally investigate intricate nanoscopic biological processes in complex living systems in real-time. The University of Exeter is renowned for its cutting-edge biomedical research, particularly in areas aligning with MRC priority areas, such as neuroscience and mental health, molecular and cellular medicine, infections and immunity, and translational research. Its world-class facilities and strong global collaborations foster scientific discoveries aimed at addressing human health challenges. To advance and keep pace with world-class research, we have identified that nano-resolution microscopy with dynamic live-imaging capabilities is currently in high demand across many research fields and departments. However, such technology is lacking on campus and is also unavailable within the UK South West research network. Therefore, we have assembled a team comprising 19 academics, an industry collaborator, a microscopy experimental officer, and a microscopy specialist. With strong support from the University of Exeter and its Bioimaging Centre, we seek to acquire the Leica TauSTED system to be housed within the Bioimaging Centre. Our aims and objectives are: To transform and elevate biomedical research at Exeter by leveraging state-of-the-art TauSTED capabilities to advance our understanding and treatment of human diseases. To bring the first TauSTED system to South West England. To enhance the existing educational platform for advanced microscopy at Exeter and within the wider community. To promote both internal and external collaborations, as well as industry partnerships. To support professional development of research and technical staff at various career stages by ensuring equal access to advanced technology. Potential applications and benefits of implementing TauSTED technology in Exeter are plentiful and include at least the following, directly related to our proposed research: Revealing dynamic nanoscopic events fundamental to neuroscience in health and disease, such as neuroglia interactions, neurodegeneration, and synaptic RNA and protein dynamics. Dissecting dynamic cell-cell and subcellular communications during embryogenesis, metabolic disorders, and diabetes at nanoscopic resolution. Understanding early immune responses and assessing drug therapies for autoimmune, heart and kidney diseases with nanoscale precision. By enabling dynamic live-imaging at nanoscale resolution, the first TauSTED system in the South West represents a major technological step forward for research and education in Exeter and its wider community. As demonstrated by the proposed applications, TauSTED promises to deliver unprecedented insights in both space and time across diverse research areas, ultimately benefiting human health by accelerating scientific discoveries in disease progression, diagnosis and treatment.