Coordinate-targeted fluorescence nanoscopy with multiple off states

2016 | journal article; research paper. A publication with affiliation to the University of Göttingen.

Jump to:Cite & Linked | Documents & Media | Details | Version history

Cite this publication

​Danzl, J. G., Sidenstein, S. C., Gregor, C., Urban, N. T., Ilgen, P., Jakobs, S. & Hell, S. W. (2016). ​Coordinate-targeted fluorescence nanoscopy with multiple off states. Nature Photonics10(2), ​122​-128​. ​doi: https://doi.org/10.1038/NPHOTON.2015.266 

Documents & Media

License

GRO License GRO License

Details

Authors
Danzl, Johann G. ; Sidenstein, Sven C. ; Gregor, Carola ; Urban, Nicolai T. ; Ilgen, Peter ; Jakobs, Stefan ; Hell, Stefan W. 
Abstract
Far-field super-resolution fluorescence microscopy discerns fluorophores residing closer than the diffraction barrier by briefly transferring them in different (typically ON and OFF) states before detection. In coordinate-targeted super-resolution variants, such as stimulated emission depletion (STED) microscopy, this state difference is created by the intensity minima and maxima of an optical pattern, causing all fluorophores to assume the off state, for instance, except at the minima. Although strong spatial confinement of the on state enables high resolution, it also subjects the fluorophores to excess intensities and state cycles at the maxima. Here, we address these issues by driving the fluorophores into a second off state that is inert to the excess light. By using reversibly switchable fluorescent proteins as labels, our approach reduces bleaching and enhances resolution and contrast in live-cell STED microscopy. Using two or more transitions to off states is a useful strategy for augmenting the power of coordinate-targeted super-resolution microscopy.
Issue Date
2016
Journal
Nature Photonics 
ISSN
1749-4885
eISSN
1749-4893
Language
English

Reference

Citations


Social Media