Lattice Selective Plane Illumination Microscopy (L-SPIM) for the analysis of subcellular dynamics in living specimens.

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The discovery of the green fluorescent protein from the jellyfish Aequorea victoria has revolutionised our way in which we study cells in a tissue but also our approaches to investigate signalling events, organelle dynamics and interactions of organelles within the complex environment of the living cell. Parallel to the developments in GFP biology, there have been advances in fluorescence imaging methods and microscopical systems that make it possible to follow fluorescently labelled cells, organelles and cytoskeleton elements, to quantify their abundance and to probe their mobility and interactions. However, long-term imaging of complex 3D tissues with conventional laser-scanning microscopes is still one of the most significant obstacles in fluorescent microscopy as this microscopy induces phototoxicity and photobleaching. Selective plane illumination microscopes (SPIM) has been developed allowing long-term imaging by scanning the specimen with much less laser-induced damage such as phototoxicity and photobleaching. However, subcellular structures deep in tissue are still a challenge to resolve. Due to the recent development of Lattice SPIM, it is now possible to image small structures such as vesicles or microtubules over an extended scanning time deep in tissue without damaging the specimen. The combined advances in GFP biology and imaging methods are providing a massive opportunity for investigating the kinetic properties of organelles in living cells. The University of Exeter studies many aspects of biological and biomedical research, reaching from fungal-related plant disease research to signalling biology in vertebrate embryonic development. In the past, microscopical approaches such as confocal microscopy and electron microscopy have been used to investigate subcellular interactions. These observations can now be complemented and extended in real-time in living cells in complex organisms. The latest generation of Lattice SPIM is, therefore, a game-changer for cell biology - in bacteria, fungi, plants and animals. It is now possible to generate dynamic maps of organelles in living cells using a fluorescence microscope over a timescale of hours to days, which has not previously been possible. In this application, we seek support to purchase a modern Lattice SPIM, which will complement the existing facilities at the Exeter Bioimaging Centre.