Understanding host interactions with high consequence pathogens using super resolution light microscopy in biocontainment.

We propose to install a state-of-the-art real time live cell imaging system in the internationally recognised Containment Level 3/outbreak response laboratories at the University of Liverpool, enhancing capabilities for live cell imaging on dangerous pathogens at the human/animal interface. We will use this equipment to: (i). Study the trafficking of proteins from infectious diseases in the cell in real time and identify the dynamic interactions with host protein complexes. (ii). Follow the effects of pathogens in real time on their host by our further development of organ slices, organoids, 3D cultures such as organ on a chip. This includes changes in morphology, the influx and efflux of different cell types in response to infection, including inflammation that is responsible for much of a disease phenotype. This focus will also allow us to contribute to the refinement, reduction and replacement in animal research. The possible gains in sensitivity (limits of detection) combined with live cell imaging at CL3, are thus unique with respect to the instrumentation, both within the University of Liverpool, and across the UK. This advanced instrumentation is essential if we are to maintain UK infectious disease research at the forefront of this rapidly developing field, which underpins mainstream biological and veterinary and biomedical research. Our exemplar projects demonstrate the breadth and broad applicability of how this advanced live cell imaging instrumentation will expand the breadth and detail of information that can be elucidated from biological samples. Specifically, we will establish the first platform in the UK for live cell imaging at high containment and improve the characterisation of the dynamic interactions that occur within a cell, tissue, and organism in response to infection.