ArticleBiophysical journal1996
Influence of the triplet excited state on the photobleaching kinetics of fluorescein in microscopy.
Article in Biophysical journal, 1996. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 68 papers.
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Who cites it
68 citing papers in PubMed, 253 citations in OpenAlex.
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- The Combined Photosensitizers in Antimicrobial Photodynamic Therapy: The Case of Methylene Blue and Photodithazine AgainstInternational journal of molecular sciences · 2025Article
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- A gentle palette of plasma membrane dyes.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Dually Labeled Neurotensin NTSJournal of medicinal chemistry · 2024Article
- Recovering true FRET efficiencies from smFRET investigations requires triplet state mitigation.Nature methods · 2024Article
- Fluorescence microscopy shadow imaging for neuroscience.Frontiers in cellular neuroscience · 2024Review
- Optical traps induce fluorophore photobleaching by two-photon excitation.Biophysical journal · 2023Article
- Turn-On Fluorescent pH Probes for Monitoring Alkaline pHs Using Bis[2-(2'-hydroxyphenyl)benzazole] Derivatives.Sensors (Basel, Switzerland) · 2023Article
- Practical Guidance for Developing Small-Molecule Optical Probes for In Vivo Imaging.Molecular imaging and biology · 2023Review
- Application of Green-enhanced Nano-lantern as a bioluminescent ratiometric indicator for measurement of Arabidopsis thaliana root apoplastic fluid pH.Plant, cell & environment · 2022Article
- Fluorescein-Based Type I Supramolecular Photosensitizer via Induction of Charge Separation by Self-Assembly.JACS Au · 2022Article
- Photosensitized Oxidation of Intracellular Targets: Understanding the Mechanisms to Improve the Efficiency of Photodynamic Therapy.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes.Biophysical reports · 2021Article
- Sources of Variability in the Response of Labeled Microspheres and B Cells during the Analysis by a Flow Cytometer.International journal of molecular sciences · 2021Article
- Photobleaching reduction in modulated super-resolution microscopy.Microscopy (Oxford, England) · 2021Article
8 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The investigation in this report aimed at providing photophysical evidence that the long-lived triplet excited state plays an important role in the non-single-exponential photobleaching kinetics of fluorescein in microscopy. Experiments demonstrated that a thiol-containing reducing agent, mercaptoethylamine (MEA or cysteamine), was the most effective, among other commonly known radical quenchers or singlet oxygen scavengers, in suppressing photobleaching of fluorescein while not reducing the fluorescence quantum yield. The protective effect against photobleaching of fluorescein in the bound state was also found in microscopy. The antibleaching effect of MEA let to a series of experiments using time-delayed fluorescence spectroscopy and nanosecond laser flash photolysis. The combined results showed that MEA directly quenched the triplet excited state and the semioxidized radical form of fluorescein without affecting the singlet excited state. The triplet lifetime of fluorescein was reduced upon adding MEA. It demonstrated that photobleaching of fluorescein in microscopy is related to the accumulation of the long-lived triplet excited state of fluorescein and that by quenching the triplet excited state and the semioxidized form of fluorescein to restore the dye molecules to the singlet ground state, photobleaching can be reduced.
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