PUBLICATION
Visualising apoptosis in live zebrafish using fluorescence lifetime imaging with optical projection tomography to map FRET biosensor activity in space and time
- Authors
- Andrews, N., Ramel, M.C., Kumar, S., Alexandrov, Y., Kelly, D.J., Warren, S.C., Kerry, L., Lockwood, N., Frolov, A., Frankel, P., Bugeon, L., McGinty, J., Dallman, M.J., French, P.M.
- ID
- ZDB-PUB-160113-7
- Date
- 2016
- Source
- Journal of biophotonics 9(4): 414-24 (Journal)
- Registered Authors
- Andrews, Natalie, Bugeon, Laurence, Dallman, Maggie, Ramel, Marie-Christine
- Keywords
- Caspase 3, FLIM, FRET, OPT, zebrafish
- MeSH Terms
-
- Optical Imaging/methods*
- Biosensing Techniques/methods*
- Proteolysis
- Spatio-Temporal Analysis
- Apoptosis*
- Caspase 3/metabolism
- Zebrafish*/metabolism
- Fluorescence Resonance Energy Transfer/methods*
- Animals
- PubMed
- 26753623 Full text @ J. Biophotonics
Citation
Andrews, N., Ramel, M.C., Kumar, S., Alexandrov, Y., Kelly, D.J., Warren, S.C., Kerry, L., Lockwood, N., Frolov, A., Frankel, P., Bugeon, L., McGinty, J., Dallman, M.J., French, P.M. (2016) Visualising apoptosis in live zebrafish using fluorescence lifetime imaging with optical projection tomography to map FRET biosensor activity in space and time. Journal of biophotonics. 9(4):414-24.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping