Controlling the fluorescence of ordinary oxazine dyes for single-molecule switching and superresolution microscopy

Vogelsang, J., Cordes, T., Forthmann, C., Steinhauer, C. & Tinnefeld, P., 2009, In : Proceedings of the National Academy of Sciences of the United States of America. 106, 20, p. 8107-8112 6 p.

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  • Jan Vogelsang
  • Thorben Cordes
  • Carsten Forthmann
  • Christian Steinhauer
  • Philip Tinnefeld
Fluorescent molecular switches have widespread potential for use as sensors, material applications in electro-optical data storages and displays, and superresolution fluorescence microscopy. We demonstrate that adjustment of fluorophore properties and environmental conditions allows the use of ordinary fluorescent dyes as efficient single-molecule switches that report sensitively on their local redox condition. Adding or removing reductant or oxidant, switches the fluorescence of oxazine dyes between stable fluorescent and non-fluorescent states. At low oxygen concentrations, the off-state that we ascribe to a radical anion is thermally stable with a lifetime in the minutes range. The molecular switches show a remarkable reliability with intriguing fatigue resistance at the single-molecule level: Depending on the switching rate, between 400 and 3,000 switching cycles are observed before irreversible photodestruction occurs. A detailed picture of the underlying photoinduced and redox reactions is elaborated. In the presence of both reductant and oxidant, continuous switching is manifested by ‘‘blinking’’ with independently controllable on- and off-state lifetimes in both deoxygenated and oxygenated environments. This ‘‘continuous switching mode’’ is advantageously used for imaging actin filament and actin filament bundles in fixed cells with subdiffraction-limited resolution.
Original languageEnglish
Pages (from-to)8107-8112
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number20
Publication statusPublished - 2009
Externally publishedYes


  • single-molecule spectroscopy, sensor, molecular switch, electron transfer, INTERFACIAL ELECTRON-TRANSFER, PHOTOCHROMISM, NANOSCOPY, SPECTROSCOPY, KINETICS, BLINKING, PROTEIN, SYSTEM, DRONPA

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