Geminate Recombination as a Photoprotection Mechanism for Fluorescent Dyes

Holzmeister, Phil; Gietl, Andreas; Tinnefeld, Philip

doi:10.1002/anie.201310300

Cited by 33 publications

(56 citation statements)

References 30 publications

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“…Apparently, TCEP uses the Cy5 triplet-state for off-switching even at very low concentrations similar to the mechanism of thiols. 28,33,36 Experiments with Cy5-COT support this notion of photo-induced off-switching via tripletstate quenching since the impact of TCEP was reduced for Cy5-COT ( Figure 4a). In Cy5-COT, both TCEP and COT apparently compete for the triplet-state and the intramolecular quencher successfully outcompetes TCEP due to its high local concentration.…”

Section: #8mentioning

confidence: 68%

On the impact of competing intra- and intermolecular triplet-state quenching on photobleaching and photoswitching kinetics of organic fluorophores

Smit

Velde

Huang

et al. 2018

Preprint

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Abstract:While buffer cocktails remain the gold-standard for photostabilization and photoswitching of fluorescent markers, intramolecular triplet-state quenchers emerge as an alternative strategy to impart fluorophores with 'self-healing' or even functional properties such as photoswitching. In this contribution, we evaluated various combinations of both approaches and show that inter-and intramolecular triplet-state quenching processes compete with each other rather than being additive or even synergistic. Often intramolecular processes dominate the photophysical situation for combinations of covalently-linked and solution-based photostabilizers and photoswitching agents. In this context we identified a new function of intramolecular photostabilizers, i.e., protection of fluorophores from reversible off-switching events caused by solution-additives, which were previously misinterpreted as photobleaching. Our studies also provide practical guidance for usage of photostabilizer-dye conjugates for STORM-type super-resolution microscopy permitting the exploitation of their improved photophysics for increased spatio-temporal resolution. Finally, we provide evidence that the biochemical environment, e.g., proximity of aromatic amino-acids such as tryptophan, reduces the photostabilization efficiency of commonly used buffer cocktails. Not only have our results important implications for a deeper mechanistic understanding of self-healing dyes, but they will provide a general framework to select label positions for optimal and reproducible photostability or photoswitching kinetics.All rights reserved. No reuse allowed without permission.(which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.

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Section: #8mentioning

confidence: 68%

On the impact of competing intra- and intermolecular triplet-state quenching on photobleaching and photoswitching kinetics of organic fluorophores

Smit

Velde

Huang

et al. 2018

Preprint

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“…According to the Marcus theory, the rate of electron transfer in the normal region increases with the thermodynamic driving force of the reaction. 40 Here, the responsiveness of a fluorophore to COT is expected to be lowered when the rate of gaining or losing electrons shortens the triplet state lifetime 9, 41 such that redox reactions are faster than the rate of triplet energy transfer with COT.…”

Section: Resultsmentioning

confidence: 99%

Intra-molecular triplet energy transfer is a general approach to improve organic fluorophore photostability

Zheng

Jockusch

Rodríguez-Calero

et al. 2016

Photochem Photobiol Sci

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Bright, long-lasting and non-phototoxic organic fluorophores are essential to the continued advancement of biological imaging. Traditional approaches towards achieving photostability, such as the removal of molecular oxygen and the use of small-molecule additives in solution, suffer from potentially toxic side effects, particularly in the context of living cells. The direct conjugation of small-molecule triplet state quenchers, such as cyclooctatetraene (COT), to organic fluorophores has the potential to bypass these issues by restoring reactive fluorophore triplet states to the ground state through intra-molecular triplet energy transfer. Such methods have enabled marked improvement in cyanine fluorophore photostability spanning the visible spectrum. However, the generality of this strategy to chemically and structurally diverse fluorophore species has yet to be examined. Here, we show that the proximal linkage of COT increases the photon yield of a diverse range of organic fluorophores widely used in biological imaging applications, demonstrating that the intra-molecular triplet energy transfer mechanism is a potentially general approach for improving organic fluorophore performance and photostability.

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“…Photoswitchable fluorescent probes have been instrumental for their unique ability of photo-conversion between a bright fluorescent state and a dark state for hundreds of cycles45. The number of photons emitted per cycle determines the effective brightness of the molecule.…”

mentioning

confidence: 99%

Reversible Photoswitching of Carbon Dots

Khan

Verma

Gupta

et al. 2015

Sci Rep

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We present a method of reversible photoswitching in carbon nanodots with red emission. A mechanism of electron transfer is proposed. The cationic dark state, formed by the exposure of red light, is revived back to the bright state with the very short exposure of blue light. Additionally, the natural on-off state of carbon dot fluorescence was tuned using an electron acceptor molecule. Our observation can make the carbon dots as an excellent candidate for the super-resolution imaging of nanoscale biomolecules within the cell.

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Geminate Recombination as a Photoprotection Mechanism for Fluorescent Dyes

Cited by 33 publications

References 30 publications

On the impact of competing intra- and intermolecular triplet-state quenching on photobleaching and photoswitching kinetics of organic fluorophores

On the impact of competing intra- and intermolecular triplet-state quenching on photobleaching and photoswitching kinetics of organic fluorophores

Intra-molecular triplet energy transfer is a general approach to improve organic fluorophore photostability

Reversible Photoswitching of Carbon Dots

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