Spinning disk microscope for Wolfson Light Microscopy Facility

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How components interact within cells is key to how cells behave and this in turn affects the organisation of tissues and ultimately the whole organism. Seeing how these interactions occur is key to our understanding of how a healthy organism forms and continues to stay healthy, and what happens when things go wrong in disease. Often visualisation of biological events causes us to formulate new hypotheses. Researchers thus use microscopy extensively to understand how cells, tissues and organisms function. Frequently, fluorescent labels are attached to components of interest which allows them to be observed in living cells. The time-scale of biological processes is highly variable: some processes happen very quickly, meaning that images need to be acquired speedily while others occur over a much longer time-scale of minutes to hours. There are a number of challenges to capturing fluorescence images because the exposure of cells to light can cause phototoxicity which damages cells and causes aberrant behaviour. Additionally, some fluorescent labels can be quite dim which makes the fluorescent signal difficult to detect.The Wolfson Light Microscopy Facility (LMF) at the University of Sheffield houses a range of microscopes that are optimised for particular applications. Spinning disk confocal microscopy allows imaging of fast dynamic events in 3 dimensions. Our current spinning disk confocal microscope is extensively used to understand how normal physiology is regulated, and what changes as organisms age. It is also used in experiments with model organisms such as zebrafish and fruit flies which help us to understand how the immune system works. Our existing system is however now greater than 12 years old and is vulnerable because the original parts are no longer supported by the manufacturers. The aim of this proposal is to install a replacement spinning disk microscope. This will not only accommodate our existing needs but will open up a range of exciting new possibilities because the new generation spinning disk confocal microscopes offer significantly enhanced performance in terms of speed, sensitivity, and low phototoxicity. These advantages, coupled with the ability to capture wide fields-of-view, mean that this is a very powerful instrument which will have a wide range of uses within the LMF. We have identified projects ranging from cell and developmental biology, immunology and microbiology which will benefit from the acquisition of a state-of-the-art spinning disk confocal microscope.