Super-resolution imaging

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Bioimaging, the investigation of cells and organisms by microscopy, is one of the key technologies for life science research. Often the way life functions has however been studied in fixed samples and by reducing cells and organisms to 2-dimensional projections. Although this approach has led and will also in the future lead to breakthrough discoveries there is a realisation that if possible "life" should be studied in a live 3D environment. Access to live cell imaging, the investigation of live cells or organisms by microscopy over time, thus is critical for state-of-the-art life science research. Live cell imaging becomes the more powerful the smaller the structures are that it can distinguish or 'resolve'. Diffraction of visible light generally limits the ability of imaging system to resolve structures to a size of 1/50th to 1/100th of a cell diameter. For a further gain in resolution a number of 'super-resolution' microscopy approaches have been developed over the last few years. Many of these approaches are however too slow to capture the rapid dynamics of life cells and organisms. A fast super-resolution approach has recently been made commercially available, called structured illumination microscopy (SIM) with lattice illumination, that is much better suited for the imaging of live cells.We apply for a state-of-the-art live cell imaging instrument with lattice SIM, the Zeiss Elyra7. Its live cell super-resolution capability is complemented by the ability to detect even smaller structures, single molecules, in fixed cells. We will integrate the technology into the Wolfson Bioimaging Facility, the centralised microscopy facility of the Faculty of Life Sciences at the University of Bristol. This facility provides access to and technical support for a large number of imaging technologies for 100s of members of staff at the University of Bristol and dozens of outside collaborators. With the Elyra7 we will, for the first time, be able to offer rapid super-resolution live cell and single molecule microscopy. To the best of our knowledge, these would be unique capabilities in the entire Southwest of the UK. The Elyra7 will be used immediately by a core group of 13 heavy users at the University of Bristol that support this application. We will implement a detailed strategy to grow use at the University, in the greater Southwest and in our user base in industry. The Elyra7 will thus enable and enhance a large number of research projects.We will apply the Elyra7 to study many research questions in live cells. These include subcellular trafficking, the movement of molecules and subcellular structures such as vesicles within a cell, and the regulation of the behaviour of various cell types that need to rapidly adapt to changing physiological conditions within our body, such as neurons and blood cells. We will also use the microscope to advance our understanding in Synthetic Biology. Some of our Synthetic Biology research focuses on the design of new protein molecules that can be used to regulate cell behaviour, for example as a platform for vaccines. Our basic research is closely linked to translational exploitation, through industrial research collaborations and a number of University spin out companies. Super-resolution microscopy will strengthen research in these collaborations and therefore generate a larger economic impact.Training of the next generation of scientists is an important aspect within the Wolfson Bioimaging Facility as it supports a large number of Ph.D. training programmes. Training in the cutting-edge live cell imaging approaches enabled by the Elyra7 will be integrated into our Ph.D. programmes. Many undergraduate students, from the University and beyond, also participate in microscopy-based research enhanced by the Elyra7. Striking images enabled by the super-resolution capabilities of the Elyra7 will strengthen outreach to secondary school students and the wider public.