SHeffield IMAging (SHIMA)

Imaging has been at the heart of huge leaps in understanding in biology and medicine, from the discovery of bacteria using the first microscopes to neuroimaging to determine brain function. All the advances have been totally dependent on the development of new technologies. This is particularly so for the study of objects too small to be seen by the naked eye, where microscopes are essential. Elucidating how cells function, whether they be from humans, plants, fungi or bacteria requires the ability to be able to see the components that make up the cell, determine their localisation and establish how they move relative to each other. In particular this has been driven by developments in microscopy coupled with the use of probes to fluorescently label molecules inside cells and so be able to localise and track them, giving key insights into processes such as cell division, growth and differentiation. However traditional optical microscopy approaches are limited in resolution to about 200 nm, which prevents the nanoscale visualisation of the myriad smaller structures and processes within the cell. Only very recently have new super-resolution (SR) microscopy approaches been invented to allow resolution down to single molecules. Thus we are at a tipping point, where existing methods and levels of understanding will become rapidly superseded by new approaches. In Sheffield we have been quick to recognise the potential of this opportunity and we are at the forefront of the development of new methods in SR microscopy. This includes building our own microscopes, coupling fluorescence microscopy with other high-resolution approaches and applying these advances to important biological and biomedical questions. We are thus ideally placed to optimally exploit the new SR technology as part of a programme of development integrated with application. The University of Sheffield (UoS) is a well-established centre with great strengths in many aspects of biomedical and biological research that are poised to reap the benefits from the new SR technologies.
We propose to establish a world-class centre in SR imaging, entitled Sheffield Imaging (SHIMA), based on our current strengths, the proposed project and a large strategic investment by the University of Sheffield. We will purchase commercial SR systems for both Structured Illumination Microscopy (SIM) and Stochastic Optical Reconstruction Microscopy (STORM) as complementary systems allowing single molecule resolution and live cell imaging. The equipment will be housed in our Light Microscopy Facility (LMF) to allow multiple user access from the outset. SR is an emerging field and so an imaging, development centre will be established to drive forward technological advances, for transfer to the LMF. To enhance technology development we will conduct specific seed projects to produce novel probes for both SIM and STORM, to increase the versatility of the techniques and to increase the speed of STORM to allow live cell imaging in 3D. With commercial partners we will ensure the rapid translation of our developments for the community. An interdisciplinary team will manage SHIMA, ensuring optimal technology development within the context of its direct application. Two SHIMA senior experimental officers (SEOs) will be responsible for interdisciplinary technology development, production of novel probes and establishment of widespread biomedical applications. The SEOs will be responsible for training of users, both internal and external creating a vibrant atmosphere of expert researchers across the University (and beyond) with a seamless interface between technology development and application via the LMF. The UoS is committed to SHIMA not only by a substantial direct investment in the project but also via a wider initiative in imaging to establish Sheffield as a key international centre in SR imaging.

Super-resolution (SR) microscopy is beginning to allow exciting new discoveries to be made across biomedical and biological disciplines. Rapid and widespread application of the new technology has been hampered by the lack of robust, affordable commercial systems and the dearth of compatible probes. We aim to establish a world-class centre in SR imaging based on our existing strengths in technology development and its biomedical application, the proposed project and a significant investment by the University of Sheffield. We will utilise commercial Structured Illumination Microscopy (SIM) and Stochastic Optical Reconstruction Microscopy (STORM) systems in a range of applications but, crucially, also embed the commercial instrumentation within a culture of technology development via a new centre for biophysical imaging. This will ensure that emerging technologies are quickly translated into wide usage by the biomedical community, further enhanced by our close association with industry, ensuring translation of our developments. At the outset we have identified probe development and increasing the image acquisition speed of STORM as key goals for our programme. The University of Sheffield will make a direct investment in the project by funding a senior experimental officer, refit of a new microscopy development facility, PhD studentships and a contribution towards the large equipment costs. The 5-year duration of the funded programme will ensure the optimal development and application of SR microscopy across the University, with greatest impact. Our ambitious project will be significantly enhanced as imaging is a flagship initiative for the University of Sheffield, creating a vibrant interface of technology to fully exploit the potential of the technologies within the context of their application to generate new levels of biomedical understanding.

The proposed programme will develop a sustainable, integrated platform of super-resolution microscopy development and its widespread application. There will be a variety of impacts over a range of timescales and arenas.

- Industry: potential users (expected timescale: year 2 onwards): will benefit by (i) being able to input into the future direction of SHIMA (ii) being able to explore potential applications for their research. The project will create a centre for integrated development and application of novel microscopy technologies.
- Industry: microscope manufacturers (expected timescale : year 4 onwards): will benefit from the technological developments arising from the initiative with potential for commercial exploitation.
- Local communities (expected timescale: year 2 onwards): via outreach activities will benefit from greater knowledge of this scientific area
- UK PLC (expected timescale year 2 onwards): will benefit through the development of imaging technologies and the commercial exploitation thereof.
- Undergraduate/postgraduate/post docs (expected timescale: year 1 onwards): will benefit through development of skills in super-resolution imaging techniques, particularly as it applies to the applications at the interface between biological and physical sciences.
- Media (expected impact: year 2 onwards): will benefit through the applicants' participation in radio and newspapers interviews.