ArticleBiology of the cell2025
Impact of triplet state population on GFP-type fluorescence and photobleaching.
Article in Biology of the cell, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- Optimizing photoactivation of PA-mCherry for optical pooled CRISPR screens.FEBS open bio · 2026Article
- All-optical strategies to minimize photobleaching in reversibly switchable fluorescent proteins.Nature communications · 2025Article
- Optimizing the in vitro neuronal microenvironment to mitigate phototoxicity in live-cell imaging.Stem cell research & therapy · 2025Article
- A fluorescent-protein spin qubit.Nature · 2025Article
- Impact of triplet state population on GFP-type fluorescence and photobleaching.Biology of the cell · 2025Article
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2 authors.
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Abstract
background informationBased on recently published parameters (Rane et al. 2023, JPCB 127, 5046-5054) for (rs)EGFP triplet state formation and decay rates and yields, we consider the power density dependence of triplet state population dynamics and its consequences for the application of green fluorescent proteins in biological single molecule fluorescence microscopy.
resultsWe find that under certain conditions, the photon budget of GFP type fluorescent proteins can be linearly dependent on power density and we propose a possible explanation for such a non-Hirschfeld photobleaching behavior. Moreover, illumination with millisecond pulses at sub-kHz rates is shown to improve photostability.
conclusionsWe stipulate that a judicious choice of excitation wavelength should take into account the triplet state absorption spectrum along with the singlet state absorption spectrum. Formulas are given for the estimation of the effects of such choice as function of the experimental parameters. SIGNIFICANCE: The linear photobleaching model as proposed by Hirschfeld 50 years ago with power-independent photon budget is not generally applicable to fluorescent proteins with millisecond-lived triplet states.
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