Fluorescence nanoscopy by polarization modulation and polarization angle narrowing
2014 | journal article; research paper. A publication with affiliation to the University of Göttingen.
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Cite this publication
Fluorescence nanoscopy by polarization modulation and polarization angle narrowing
Hafi, N.; Grunwald, M. ; van den Heuvel, L. S.; Aspelmeier, T. ; Chen, J.-H. ; Zagrebelsky, M. & Schütte, O. M. et al. (2014)
Nature Methods, 11(5) pp. 579-584. DOI: https://doi.org/10.1038/NMETH.2919
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Details
- Authors
- Hafi, Nour; Grunwald, Matthias ; van den Heuvel, Laura S.; Aspelmeier, Timo ; Chen, Jian-Hua ; Zagrebelsky, Marta; Schütte, Ole M. ; Steinem, Claudia ; Korte, Martin; Munk, Axel ; Walla, Peter J.
- Abstract
- When excited with rotating linear polarized light, differently oriented fluorescent dyes emit periodic signals peaking at different times. We show that measurement of the average orientation of fluorescent dyes attached to rigid sample structures mapped to regularly defined (50 nm)(2) image nanoareas can provide subdiffraction resolution (super resolution by polarization demodulation, SPoD). Because the polarization angle range for effective excitation of an oriented molecule is rather broad and unspecific, we narrowed this range by simultaneous irradiation with a second, de-excitation, beam possessing a polarization perpendicular to the excitation beam (excitation polarization angle narrowing, ExPAN). This shortened the periodic emission flashes, allowing better discrimination between molecules or nanoareas. Our method requires neither the generation of nanometric interference structures nor the use of switchable or blinking fluorescent probes. We applied the method to standard wide-field microscopy with camera detection and to two-photon scanning microscopy, imaging the fine structural details of neuronal spines.
- Issue Date
- 2014
- Journal
- Nature Methods
- ISSN
- 1548-7091
- eISSN
- 1548-7105
- Language
- English