The University of Bath Multi-user Light Sheet Fluorescence Microscopy

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We wish to add an important cutting-edge instrument to the University of Bath's microscopy facilities. Many aspects of physiology, cell and developmental biology are dynamic, including those on second (e.g. heart-beat), minute (e.g. cell migration) and hour or even day timescales (e.g. embryonic development and plant growth). Likewise, biological systems are frequently 3-dimensional, with structural arrangements being integral to their function (e.g. cardiovascular systems, brain, embryos). Thus, to study biological processes in detail, we need microscopy that can allow us to image biological processes on this range of timescales, and in intact 3-D samples (whole organisms, or tissues). A new technique, Light Sheet Fluorescent Microscopy (LSFM), allows all of these and promises to revolutionise these areas. Key aspects of the LSFM design ensure fast imaging, so that changes on time-scales of seconds-minutes can be resolved; gentle imaging, so that specimens are not damaged over long-term study for minutes, hours or even days; and deeper imaging, so that thicker specimens and whole living organisms become accessible. It means that we can study changes in response to drugs or pathogens over long timescales; that we can monitor cells as they migrate through complex tissues or even whole organisms, detecting changes in their behaviour as their environment alters; and that we can study processes in living cells organised into 3-D structures that more accurately mimic a physiological environment. In Bath, we propose using the LSFM for seven initial projects, each utilising one or more key features of LSFM: to study stem cells in developing zebrafish embryos, fat development in mouse models of body mass control, cell environment on cartilage properties, how bacterial communities (biofilms) become more efficient at evading antibiotic treatment, plant root responses to infection, how to make reproducible organoid cultures, and mapping neuronal activation in mouse brains. The instrument will be integrated into the Microscopy and Analysis Suite in the University of Bath, ensuring expert user training and equipment support. It will support a broad, interdisciplinary research community within the University of Bath and our collaborator institutions/organisations.