DOI: 10.1073/pnas.0811875106. Controlling the fluorescence of ordinary oxazine dyes for single-molecule switching and superresolution microscopy

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

10.1073/pnas.0811875106

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

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 nonfluorescent 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.

Bibliography

Vogelsang, J., Cordes, T., Forthmann, C., Steinhauer, C., & Tinnefeld, P. (2009). Controlling the fluorescence of ordinary oxazine dyes for single-molecule switching and superresolution microscopy. Proceedings of the National Academy of Sciences, 106(20), 8107â8112.

Authors 5

Jan Vogelsang (first)
Thorben Cordes (additional)
Carsten Forthmann (additional)
Christian Steinhauer (additional)
Philip Tinnefeld (additional)

References 42 Referenced 251

BL Feringa Molecular Switches (Wiley-VCH, Weinheim, Germany), pp. 454 (2001). (10.1002/3527600329) / Molecular Switches by Feringa BL (2001)
10.1126/science.1137395
10.1002/anie.200803246
10.1021/ja0343104
10.1073/pnas.0406119101
10.1021/cr980068l
10.1038/41048
10.1073/pnas.0500489102
10.1021/ja044686x
10.1103/PhysRevLett.94.108101
10.1038/420759a
10.1021/ja047169n
10.1021/ja069131b
10.1021/ja047386o
10.1021/ja0268074
10.1002/(SICI)1438-5171(200004)1:1<17::AID-SIMO17>3.0.CO;2-E
10.1038/373132a0
10.1021/jp9634518
10.1021/jp034906d
10.1002/anie.200790129
10.1021/jp034723r
10.1038/nmeth934
10.1002/anie.200801518
10.1038/nmeth.1257
10.1021/ja806590m
10.1529/biophysj.104.045971
10.1529/biophysj.107.117689
AF Holleman, E Wiberg Lehrbuch der Anorganischen Chemie (de Gruyter, New York, 1995). / Lehrbuch der Anorganischen Chemie by Holleman AF (1995)
10.1021/ar0401348
10.1073/pnas.0700629104
10.1002/1361-6374(199609)4:3<149::AID-BIO5>3.0.CO;2-D
10.1002/ijch.197000029
10.1063/1.1742723
10.1063/1.2034116
10.1126/science.1127344
10.1038/nmeth929
10.1529/biophysj.106.091116
10.1007/s00340-008-3250-9
10.1007/s00340-007-2729-0
10.1016/S0006-3495(02)75618-X
10.1002/cphc.200800299
10.1021/ja809117z

Dates

Type	When
Created	16 years, 3 months ago (May 11, 2009, 9:43 p.m.)
Deposited	3 years, 4 months ago (April 12, 2022, 5:37 p.m.)
Indexed	3 weeks, 5 days ago (Aug. 7, 2025, 4:29 p.m.)
Issued	16 years, 3 months ago (May 19, 2009)
Published	16 years, 3 months ago (May 19, 2009)
Published Online	16 years, 3 months ago (May 19, 2009)
Published Print	16 years, 3 months ago (May 19, 2009)

Funders 0

None

BibTeX

@article{Vogelsang_2009, title={Controlling the fluorescence of ordinary oxazine dyes for single-molecule switching and superresolution microscopy}, volume={106}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0811875106}, DOI={10.1073/pnas.0811875106}, number={20}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Vogelsang, Jan and Cordes, Thorben and Forthmann, Carsten and Steinhauer, Christian and Tinnefeld, Philip}, year={2009}, month=may, pages={8107–8112} }

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