The long-awaited FAST tags for multiplex FLIM (Fluorescence Lifetime Imaging Microscopy) are now here! The team led by Arnaud Gautier of Sorbonne University indeed disclosed FAST-based fluorescence lifetime-modulating tags days ago in Adv. Sci. 2024. Those FAST variants enable tuning the fluorescence lifetime of their cognate fluorogen. As a result they allow imaging various proteins of interest using the same fluorogen and only one channel! Almost simultaneously, the team led by Mikhail S. Baranov of the Russian Academy of Sciences in Moscow released a paper in Commun. Biol. 2024 around the same matters with different FAST variants and fluorogens!
Multiplexed FLIM with FAST – © 2024 The Authors (in Adv. Sci.)
FLIM opens new dimensions for highly multiplexed imaging in live cells and organisms using differences in fluorescence lifetime to distinguish spectrally identical fluorescent probes. Both teams describe sets of three FAST tags capable of modulating the fluorescence lifetime of bound fluorogenic HBR derivatives. They show that changes in the FAST protein sequence can vary the local environment of the chromophore and lead to significant changes in its fluorescence lifetime. This means that the same fluorophore will emit light at different times, making it possible to differentiate between the labeled proteins based on when fluorescence is detected rather than the color of the fluorophore.
“Beyond multiplexed imaging, fluorescence lifetime-modulating tags could also be used for the design of lifetime responsive biosensors through coupling with recognition domains of specific analytes and biomolecules of interest. We believe fluorescence lifetime multiplexing using chemogenetic fluorescence lifetime-modulating tags opens exciting prospects for biological imaging: combined with spectral multiplexing, it opens countless opportunities for observing several biological targets simultaneously”
Prof. Arnaud Gautier
Read more about multiplex FLIM with FAST
, , , , , , , , Multiplexed In Vivo Imaging with Fluorescence Lifetime-Modulating Tags. Adv. Sci. 2024, 2404354. https://doi.org/10.1002/advs.202404354
Bogdanova, Y. A., Solovyev, I. D., Baleeva, N. S., Myasnyanko, I. N., Gorshkova, A. A., Gorbachev, D. A., … & Baranov, M. S. Fluorescence lifetime multiplexing with fluorogen activating protein FAST variants. Communications Biology, 7(1), 799. https://doi.org/10.1038/s42003-024-06501-1
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