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Abstract
We perform wide-field microscopy to investigate the photobleaching of organic fluorophores embedded in the polymeric host PMMA. Our experimental arrangement facilitates the comparison between the ensemble and single molecule data. We characterize the photostability of dye molecules of various families by measuring the ‘bleaching number’, defined as the average number of photons a molecule emits until photobleaching occurs. In particular, we have analysed the dependence of the bleaching number on the presence of oxygen. Surprisingly, we find an improvement of photostability in the presence of oxygen for ionic dyes (DiI, TMR, Rh6G, Alexa 546), suggesting that oxygen quenches the photoactive triplet state, but it only indirectly contributes to photochemistry. In contrast, we observe that photobleaching of the aromatic hydrocarbon is strongly enhanced by oxygen.
Acknowledgements
We would like to thank Dr Gert Zumofen for helpful discussions. This work was supported by ETH Zurich, the Swiss National Science Foundation (SNF) and the Swiss Ministry of Science and Education (EU Integrated Project ‘Molecular Imaging’).
