Mechanistic investigation of mEos4b reveals a strategy to reduce track interruptions in sptPALM

De Zitter, E.; Thedie, D.; Monkemoller, V.; Hugelier, S.; Beaudouin, J.; Adam, V.; Byrdin, M.; Van Meervelt, L.; Dedecker, P.; Bourgeois, D.

doi:10.1101/475939

bioRxiv · 10.1101/475939

Mechanistic investigation of mEos4b reveals a strategy to reduce track interruptions in sptPALM

De Zitter, E.·Thedie, D.·Monkemoller, V.·Hugelier, S.·Beaudouin, J.·Adam, V.·Byrdin, M.·Van Meervelt, L.·Dedecker, P.·Bourgeois, D.

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Abstract

Green-to-red photoconvertible fluorescent proteins repeatedly enter dark states, causing interrupted tracks in single-particle-tracking localization microscopy (sptPALM). We identified a long-lived dark state in photoconverted mEos4b that results from isomerization of the chromophore and efficiently absorbs cyan light. Addition of weak 488-nm light swiftly reverts this dark state to the fluorescent state. This strategy largely eliminates slow blinking and enables the recording of significantly longer tracks in sptPALM with minimum effort.

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De Zitter, E., Thedie, D., Monkemoller, V., Hugelier, S., Beaudouin, J., Adam, V., Byrdin, M., Van Meervelt, L., Dedecker, P., Bourgeois, D.. 2018-11-21. Mechanistic investigation of mEos4b reveals a strategy to reduce track interruptions in sptPALM. https://doi.org/10.1101/475939

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