A multi-disciplinary proposal for a new confocal microscope at the Royal Veterinary College

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This is a multi-disciplinary proposal to enable the Royal Veterinary College (RVC) to obtain a new confocal laser-scanning microscope (CLSM). The 100 strong research staff at the RVC gained a score of 5 in the last RAE. Their interests are encompassed by areas of research focus, three of which are represented here: musculoskeletal infection and immunity, cardiovascular biology and infection. Much of the pioneering and competitive work in these areas is done at the microscopic and molecular level. However, at a time when, more than ever, it is important to demonstrate high quality images in support of cutting-edge science, the RVC is hampered by possession of only a single CLSM that is nearly ten years old and which lacks features that our international competitors take for granted. For example, it is not possible to excite fluorophores in the near-UV, therefore the use of dyes such as DAPI and Hoechst, and techniques such as FRET and FRAP, are currently not possible. In any case, current CLSM use, even for red/green dual emission, is so high that user time is restricted. The instrument that we request is a Leica SP2 RS AOBS CLSM. This microscope is a four-channel instrument (three channel spectral detection plus transmitted light) and is fitted with four visible lasers and a 405 nm blue diode laser capable of exciting DAPI and Hoechst dyes (needed in all 10 projects). It is replete with three-dimensional and time-based software (required by projects 1, 3 and 5), and we have requested a cell incubation chamber and CCD camera to ensure live cell CLS microscopy and capture of low intensity emission (required by projects 1-3 and 5-8). This microscope will also facilitate advanced techniques such as FRET (project 1) and FRAP (project 7). In short, the CLSM requested will satisfy the research needs of all the applicants. The multi-disciplinary dimension of this proposal is evident from the individual projects, which investigate aspects of neuronal movement during development (projects 1 and 6), damaging effects on vasculature during development (project 8), trauma (project 3) and disease (project 5), functional aspects of the musculoskeletal system including satellite cell activation during muscle development (project 4), damage accumulation in injury-prone tendons (project 7) and the adaptive control of bone strength (project 10), aspects of aging and immunity (project 9) and the mechanism of prion disease (project 2). The Leica RS CLSM will facilitate progress in all of these projects; furthermore, as a key component of the physical research infrastructure, the CLSM will be available to all researchers on either campus of the RVC to further all research endeavours. All projects described here fall within the BBSRC Strategic Plan.