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CoMPLEX PhD Student Publication
24 October 2019
Visualising apoptosis in live zebrafish using fluorescencelifetime imaging with optical projection tomographyto map FRET biosensor activity in space and time. Third year PhD student, Nicola Lockwood has had another paper published in the Journal of Biophotonics.
Fluorescence lifetime imaging (FLIM) combined with op-tical projection tomography (OPT) has the potential tomap Förster resonant energy transfer (FRET) readouts inspace and time in intact transparent or near transparentlive organisms such as zebrafish larvae, thereby providinga means to visualise cell signalling processes in their phy-siological context. Here the first application of FLIMOPT to read out biological function in live transgenic zeb-rafish larvae using a genetically expressed FRET biosen-sor is reported. Apoptosis, or programmed cell death, ismapped in 3-D by imaging the activity of a FRET biosen-sor that is cleaved by Caspase 3, which is a key effector ofapoptosis. Although apoptosis is a naturally occurringprocess during development, it can also be triggered in avariety of ways, including through gamma irradiation.FLIM OPT is shown here to enable apoptosis to be mon-itored over time, in live zebrafish larvae via changes inCaspase 3 activation following gamma irradiation at24 hours post fertilisation. Significant apoptosis was ob-served at 3.5 hours post irradiation, predominantly in thehead region.