Intrinsically Resolution Enhancing Probes for Confocal Microscopy

Vogelsang, J., Cordes, T., Forthmann, C., Steinhauer, C. & Tinnefeld, P., 2010, In : Nano Letters. 10, 2, p. 672-679 8 p.

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  • Jan Vogelsang
  • Thorben Cordes
  • Carsten Forthmann
  • Christian Steinhauer
  • Philip Tinnefeld
In recent years different implementations of superresolution microscopy based on targeted switching (STED, GSD, and SSIM) have been demonstrated. The key elements to break the diffraction barrier are two distinct molecular states that generate a saturable nonlinear fluorescence response with respect to the excitation intensity. In this paper, we demonstrate that a nonlinearity can even be encoded in fluorescent probes, which then increase the resolution of a standard confocal microscope. This nonlinearity is achieved by an intensity dependent blocking of the resonance energy transfer between a donor and one or more acceptor fluorophores, utilizing radical anion states of the acceptor. In proof-of-principle experiments, we demonstrate a significant resolution increase using probes with different numbers of acceptor fluorophores. Quantitative description by a theoretical model paves the way for the development of fluorescent probes that can more than double the resolution of essentially any confocal microscope in all three dimensions.
Original languageEnglish
Pages (from-to)672-679
Number of pages8
JournalNano Letters
Issue number2
Publication statusPublished - 2010


  • nonlinear fluorescence, excited-state saturation, single-molecule spectroscopy, FRET, superresolution microscopy

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