Intramolecular TT-energy transfer in bifluorophoric laser dyes

Schäfer, F. P.; Zhang, F. G.; Jethwa, J.

doi:10.1007/bf00693890

Cited by 17 publications

(13 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The strategy to label GFP on the outside of the ß-barrel may reduce efficient interaction between the chromophore and the photostabilizer. While, there is convincing published evidence that the ß-barrel does not shield the FPchromophore fully [51][52][53] from interacting molecules in the buffer and also that tripletquenching proesses might be mediated by a contact-less Förster mechanisms 50 , we speculate that selecting a residue inside the ß-barrel might be even more promising.…”

Section: Summary and Discussionmentioning

confidence: 75%

“…While the latter are believed to occur more likely via Dexter-processes [22][23] , which would require collisions between FP chromophore and photostabilizer, there is support that certain triplet quenchers might utilize a Förster mechanism 50 . We thus reasoned that intramolecular triplet-quenching in FPs might not strictly require direct contacts between chromophore and stabilizer but proximity.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Front Cover: Characterization of Fluorescent Proteins with Intramolecular Photostabilization**

et al. 2021

View full text Add to dashboard Cite

Genetically encodable fluorescent proteins have revolutionized biological imaging in vivo and in vitro. Since there are no other natural fluorescent tags with comparable features, the impact of fluorescent proteins for biological research cannot be overemphasized. Despite their importance, their photophysical properties, i.e., brightness, count-rate and photostability, are relatively poor compared to synthetic organic fluorophores or quantum dots. Intramolecular photostabilizers were recently rediscovered as an effective approach to improve photophysical properties. The approach uses direct conjugation of photostablizing compounds such as tripletstate quenchers or redox-active substances to an organic fluorophore, thereby creating high local concentrations of photostabilizer. Here, we introduce an experimental strategy to screen for the effects of covalently-linked photostabilizers on fluorescent proteins. We recombinantly produced a double cysteine mutant (A206C/L221C) of -GFP for attachment of photostabilizermaleimides on the ß-barrel in close proximity to the chromophore. Whereas labelling with photostabilizers such as Trolox, Nitrophenyl, and Cyclooctatetraene, which are often used for organic fluorophores, had no effect on -GFP-photostability, a substantial increase of photostability was found upon conjugation of -GFP to an azobenzene derivative. Although the mechanism of the photostabilizing effects remains to be elucidated, we speculate that the higher triplet-energy of azobenzene might be crucial for triplet-quenching of fluorophores in the near-UV and blue spectral range. Our study paves the way towards the development and design of a second generation of fluorescent proteins with photostabilizers placed directly in the protein barrel by methods such as unnatural amino acid incorporation.

show abstract

Section: Summary and Discussionmentioning

confidence: 75%

Section: Resultsmentioning

confidence: 99%

Front Cover: Characterization of Fluorescent Proteins with Intramolecular Photostabilization**

et al. 2021

View full text Add to dashboard Cite

show abstract

“…History, Development, and State of the Art of Self-Healing Dyes . The concept of enhancing the photostability of fluorescent dyes via linking triplet state-quenchers was proposed, experimentally realized, and verified in the early 1980s. ,, Lüttke and co-workers linked 1,4-bis(5-phenyloxazol-2-yl)benzene (POPOP) and its derivatives to stilbene with the goal of quenching triplet states (Figure a). These self-healing POPOP–stilbene constructs showed very good performance as lasing medium in dye lasers (Figure a) and were the first self-healing dyes with intramolecular photostabilization …”

mentioning

confidence: 99%

“…Dye 2 has two CH 2 groups, whereas dye 3 has one CH 2 group between DMP and trans -stilbene. Oscillograms are taken from ref . (c) Measured triplet lifetimes τ T and calculated TT energy-transfer rates k ET .…”

mentioning

confidence: 99%

See 1 more Smart Citation

Self-Healing Dyes—Keeping the Promise?

Isselstein

Zhang

Glembockyte

et al. 2020

J. Phys. Chem. Lett.

View full text Add to dashboard Cite

Self-healing dyes have emerged as a new promising class of fluorescent labels. They consist of two units, a fluorescent dye and a photostabilizer. The latter heals whenever the fluorescent dye is in danger of taking a reaction pathway toward photobleaching. We describe the underlying concepts and summarize the developmental history and state-of-the-art, including latest applications in high-resolution microscopy, live-cell, and single-molecule imaging. We further discuss remaining limitations, which are (i) lower photostabilization of most self-healing dyes when compared to solution additives, (ii) limited mechanistic understanding on the influence of the biochemical environment and molecular oxygen on self-healing, and (iii) the lack of cheap and facile bioconjugation strategies. Finally, we provide ideas on how to further advance self-healing dyes, show new data on redox blinking caused by double-stranded DNA, and highlight forthcoming work on intramolecular photostabilization of fluorescent proteins.

show abstract

Cis‐TransPhotoisomerization of Stilbenes and Stilbene‐Like Molecules

Görner¹,

Kuhn²

1994

Advances in Photochemistry

View full text Add to dashboard Cite

Intramolecular TT-energy transfer in bifluorophoric laser dyes

Cited by 17 publications

References 16 publications

Front Cover: Characterization of Fluorescent Proteins with Intramolecular Photostabilization**

Front Cover: Characterization of Fluorescent Proteins with Intramolecular Photostabilization**

Self-Healing Dyes—Keeping the Promise?

Cis‐TransPhotoisomerization of Stilbenes and Stilbene‐Like Molecules

Contact Info

Product

Resources

About