This project aims to develop and apply a new optical tomographic imaging platform to directly observe the progression of disease, including quantitative assays of morphological changes, physiological and molecular signalling processes in live whole juvenile and adult zebrafish that benefit from fully developed vasculature and immune system. We aim to extend optical projection tomography (OPT) to rapid imaging of cell migration using a novel (patent filed) configuration and realise 3-D mapping of optical absorption and fluorescence signals (including fluorescence lifetime imaging - FLIM) in cm-scale non-pigmented zebrafish lines using novel computational reconstruction algorithms to account for weakly scattered light propagation and ballistic light techniques to improve 3-D image quality. We will develop this technology in the context of its application to disease models for both cancer and inflammation. For this we will perform crosses and modify already available zebrafish strains, with fluorescently labelled vasculature and cell types (tumour cells, neutrophils and macrophages). For cancer we would make 3-D measurements of developing (inducible, syngeneic) tumours and the surrounding vasculature and map the localisation and size of resulting metastases over time. We would also read out the activation of Src, MT1-MMP & Rac biosensors using FLIM FRET and correlate changes in these important signalling pathways with tumour development. For inflammation we would image the kinetics of recruitment of inflammatory cell populations throughout a single fish in response to chemical pollutants and correlate this with the activation of signalling networks (Nalp3 inflammasome, TLR) using sensors for cathepsin B, caspase 1 and NF-kB. We would thereby study how chemically induced inflammation varies between larval, juvenile and adult zebrafish. As a basis for future projects, we would start exploring the inflammatory response in tumour bearing zebrafish.