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Absolute quantum yield measurements of fluorescent proteins using a plasmonic nanocavity.


ABSTRACT: One of the key photophysical properties of fluorescent proteins that is most difficult to measure is the quantum yield. It describes how efficiently a fluorophore converts absorbed light into fluorescence. Its measurement using conventional methods become particularly problematic when it is unknown how many of the proposedly fluorescent molecules of a sample are indeed fluorescent (for example due to incomplete maturation, or the presence of photophysical dark states). Here, we use a plasmonic nanocavity-based method to measure absolute quantum yield values of commonly used fluorescent proteins. The method is calibration-free, does not require knowledge about maturation or potential dark states, and works on minute amounts of sample. The insensitivity of the nanocavity-based method to the presence of non-luminescent species allowed us to measure precisely the quantum yield of photo-switchable proteins in their on-state and to analyze the origin of the residual fluorescence of protein ensembles switched to the dark state.

SUBMITTER: Ruhlandt D 

PROVIDER: S-EPMC7599333 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Absolute quantum yield measurements of fluorescent proteins using a plasmonic nanocavity.

Ruhlandt Daja D   Andresen Martin M   Jensen Nickels N   Gregor Ingo I   Jakobs Stefan S   Enderlein Jörg J   Chizhik Alexey I AI  

Communications biology 20201030 1


One of the key photophysical properties of fluorescent proteins that is most difficult to measure is the quantum yield. It describes how efficiently a fluorophore converts absorbed light into fluorescence. Its measurement using conventional methods become particularly problematic when it is unknown how many of the proposedly fluorescent molecules of a sample are indeed fluorescent (for example due to incomplete maturation, or the presence of photophysical dark states). Here, we use a plasmonic n  ...[more]

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2024-09-30 | GSE271876 | GEO