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Quenching of Long Lifetime Emitting Fluorophores with Paramagnetic Molecules

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Abstract

In this work, we have studied quenching of the fluorescence of two well-known oxygen probes, 1-pyrene butyric acid (PBA) and tris(2,2’-bipyridine)ruthenium ([Ru(bpy)3]2+) by reactive oxygen species (superoxide anion, nitric oxide derivative, hydrogen peroxide) and by the O2 molecule. Both, time-resolved and steady state fluorescence measurements were performed in solution (ethanol, dimethyl sufoxide, water) and in micelles of Sodium Dodecyl Sulfate that serve as a model for membrane-containing biological structures. We have found that only the free radicals and O2 can actively quench for the two probes, but not the diamagnetic H2O2. Our data correspond to the classical Stern–Volmer equation. H2O2 has an effect only at high molar concentrations (>0.1 M). In contrast, effective concentrations of free radicals and O2 that lead to quenching are in millimolar range. In conclusion, our methods allows for detecting global ROS that are small free radicals without interference from the reactive hydroxyl radical. Our data suggest that the method can be used for the quantification of ROS in individual living cells based on the measurement of fluorescence lifetime of those probes.

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Acknowledgments

This work was financially supported by the French “Ligue Contre le Cancer- Comité départemental du Gard”.

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Authors and Affiliations

  1. Department of Chemistry, University of Dokuz Eylul, Tinaztepe, Buca, Izmir, Turkey

    O. Oter

  2. Institut de Modélisation et d’Analyse en Géo-Environnements et Santé, Université de Perpignan Via Domitia, Perpignan, France

    A-C. Ribou

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  1. O. Oter
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Oter, O., Ribou, AC. Quenching of Long Lifetime Emitting Fluorophores with Paramagnetic Molecules. J Fluoresc 19, 389–397 (2009). https://doi.org/10.1007/s10895-008-0425-z

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