title: Visualising apoptosis in live zebrafish using fluorescence lifetime imaging with optical projection tomography to map FRET biosensor activity in space and time.
abstract: Fluorescence lifetime imaging (FLIM) combined with optical projection tomography (OPT) has the potential to map Förster resonant energy transfer (FRET) readouts in space and time in intact transparent or near transparent live organisms such as zebrafish larvae, thereby providing a means to visualise cell signalling processes in their physiological context. Here the first application of FLIM OPT to read out biological function in live transgenic zebrafish larvae using a genetically expressed FRET biosensor is reported. Apoptosis, or programmed cell death, is mapped in 3-D by imaging the activity of a FRET biosensor that is cleaved by Caspase 3, which is a key effector of apoptosis. Although apoptosis is a naturally occurring process during development, it can also be triggered in a variety of ways, including through gamma irradiation. FLIM OPT is shown here to enable apoptosis to be monitored over time, in live zebrafish larvae via changes in Caspase 3 activation following gamma irradiation at 24 hours post fertilisation. Significant apoptosis was observed at 3.5 hours post irradiation, predominantly in the head region.
authors.affiliation: Institute of Chemical Biology, Department of Chemistry, Imperial College London, SW7 2AZ, UK.|Department of Life Sciences, , Imperial College London, SW7 2AZ, UK.|Photonics Group, Department of Physics, Prince Consort Road, Imperial College London, SW7 2AZ, UK.
authors.forename: Natalie
authors.initials: N
authors.lastname: Andrews
authors.affiliation: Department of Life Sciences, , Imperial College London, SW7 2AZ, UK.|Division of Medicine, University College London, Gower Street, London, WC1E 6BT, UK.
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Ramel
authors.affiliation: Photonics Group, Department of Physics, Prince Consort Road, Imperial College London, SW7 2AZ, UK.
authors.forename: Sunil
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Photonics Group, Department of Physics, Prince Consort Road, Imperial College London, SW7 2AZ, UK.
authors.forename: Yuriy
authors.initials: Y
authors.lastname: Alexandrov
authors.affiliation: Photonics Group, Department of Physics, Prince Consort Road, Imperial College London, SW7 2AZ, UK.
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Kelly
authors.affiliation: Photonics Group, Department of Physics, Prince Consort Road, Imperial College London, SW7 2AZ, UK.
authors.forename: Sean C
authors.initials: SC
authors.lastname: Warren
authors.affiliation: Department of Life Sciences, , Imperial College London, SW7 2AZ, UK.
authors.forename: Louise
authors.initials: L
authors.lastname: Kerry
authors.affiliation: Division of Medicine, University College London, Gower Street, London, WC1E 6BT, UK.|COMPLEX, University College London, Gower Street, London, WC1E 6BT, UK.
authors.forename: Nicola
authors.initials: N
authors.lastname: Lockwood
authors.affiliation: Division of Medicine, University College London, Gower Street, London, WC1E 6BT, UK.
authors.forename: Antonina
authors.initials: A
authors.lastname: Frolov
authors.affiliation: Division of Medicine, University College London, Gower Street, London, WC1E 6BT, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Frankel
authors.affiliation: Department of Life Sciences, , Imperial College London, SW7 2AZ, UK.
authors.forename: Laurence
authors.initials: L
authors.lastname: Bugeon
authors.affiliation: Photonics Group, Department of Physics, Prince Consort Road, Imperial College London, SW7 2AZ, UK.
authors.forename: James
authors.initials: J
authors.lastname: McGinty
authors.affiliation: Department of Life Sciences, , Imperial College London, SW7 2AZ, UK. m.dallman@imperial.ac.uk.
authors.forename: Margaret J
authors.initials: MJ
authors.lastname: Dallman
authors.affiliation: Photonics Group, Department of Physics, Prince Consort Road, Imperial College London, SW7 2AZ, UK. paul.french@imperial.ac.uk.
authors.forename: Paul M W
authors.initials: PM
authors.lastname: French
mesh_terms: D000818:Animals; D017209:Apoptosis; D015374:Biosensing Techniques / Q000379:methods*; D053148:Caspase 3 / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*; D059748:Proteolysis; D062211:Spatio-Temporal Analysis; D015027:Zebrafish / Q000378:metabolism
chemical_list: D053148:Caspase 3
keywords: Caspase 3; FLIM; FRET; OPT; zebrafish