A Bessel beam light sheet microscope

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Fluorescence microscopy has become a very important technique in the biosciences because it allows the distributions of specific proteins to be imaged. The resolution it provides is not as good as electron microscopy, but it has the major advantage that it can be used in live cells. However, the imaging process can still have a toxic effect on cells, and this is a major limitation in cell biology studies.A popular way to minimise the effects of imaging on cells is called light sheet microscopy. This only illuminates the part of the sample that is in focus, meaning many more pictures can be taken without damaging the cell. Generally, this also makes the resolution worse - but an approach has recently been developed which not only illuminates using a light sheet, but also improves the resolution. This is achieved using Bessel beams, which use interference and are therefore not subject to the same limitations as most microscopy techniques. Using this lattice light sheet approach, the sample can be imaged quickly and with excellent resolution.We will build a replica of the lattice light sheet system at Janelia Research Campus (where the technique was developed) in the Microscopy Innovation Centre (MIC) at King's College London, using blueprints and equipment lists provided to us by Janelia. The MIC has been created by KCL in recognition of the central role that microscopy plays in biological and biomedical innovation. It's purpose is to make advanced microscopy techniques available to users, and this is made possible by two experienced optical microscopy specialists. They will build the system, maintain it, and make it available to users. A particularly notable feature of this system is that it will have an extra module to allow specific areas of the samples to either be bleached or for fluorophores which can emit in two different colours to be switched from one to the other. We believe this would be the first system of its kind in the UK.A wide range of different project applications have been proposed. These include three from groups who already travel to Janelia Research Campus to use the system there, so they are familiar with the equipment and sample preparation requirements. Projects will improve understanding of various basic biological processes: cell migration, cell division, how cancer cells invade, the characteristics of neurons which have been derived from stem cells, and the cytoskeletal structure and mitochondria in heart cells. The lattice light sheet microscope will let us image these live systems at very high resolution, and observe how they change over time, enabling new biological discoveries. After an initial evaluation period where the microscope is only available to users at King's, the microscope will be opened to users in other institutions, who would them be able to access this system with full technical support without travelling to the USA. The ability to image cells at high resolution, without the the imaging process changing their behaviour, promises to vastly increase the amount of information available about cells. By making so much more information available, this microscope will help to maintain the UK's leading position in cell biology and biomedical research.