How many photons are needed for FRET imaging?
Biomedical optics express
The Optical Society
MetadataShow full item record
Esposito, A. (2020). How many photons are needed for FRET imaging?. Biomedical optics express, 11 (2), 1186-1202. https://doi.org/10.1364/boe.379305
FRET imaging is an essential analytical method in biomedical research. The limited photon-budget experimentally available, however, imposes compromises between spatiotemporal and biochemical resolutions, photodamage and phototoxicity. The study of photon-statistics in biochemical imaging is thus important in guiding the efficient design of instrumentation and assays. Here, we show a comparative analysis of photon-statistics in FRET imaging demonstrating how the precision of FRET imaging varies vastly with imaging parameters. Therefore, we provide analytical and numerical tools for assay optimization. FLIM is a very robust technique with excellent photon-efficiencies but also intensity-based FRET imaging can reach very high precision by utilizing also information within acceptor fluorescence.
MRC (MC_UU_12022/1_do not transfer?)
Medical Research Council (MC_UU_12022/1)
External DOI: https://doi.org/10.1364/boe.379305
This record's URL: https://www.repository.cam.ac.uk/handle/1810/301310
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