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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editors: Andrew Dunn and Anthony Durkin
  • Vol. 8, Iss. 2 — Mar. 4, 2013

Frequency dependent detection in a STED microscope using modulated excitation light

Emiliano Ronzitti, Benjamin Harke, and Alberto Diaspro  »View Author Affiliations

Optics Express, Vol. 21, Issue 1, pp. 210-219 (2013)

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We present a novel concept adaptable to any kind of STED microscope in order to expand the limited number of compatible dyes for performing super resolution imaging. The approach is based on an intensity modulated excitation beam in combination with a frequency dependent detection in the form of a standard lock-in amplifier. This enables to unmix fluorescence signal originated by the excitation beam from the fluorescence caused by the STED beam. The benefit of this concept is demonstrated by imaging biological samples as well as fluorescent spheres, whose spectrum does not allow STED imaging in the conventional way. Our concept is suitable with CW or pulsed STED microscope and can thereby be seen as a general improvement adaptable to any existing setup.

© 2013 OSA

OCIS Codes
(180.0180) Microscopy : Microscopy
(180.1790) Microscopy : Confocal microscopy

ToC Category:

Original Manuscript: November 12, 2012
Revised Manuscript: December 20, 2012
Manuscript Accepted: December 21, 2012
Published: January 3, 2013

Virtual Issues
Vol. 8, Iss. 2 Virtual Journal for Biomedical Optics

Emiliano Ronzitti, Benjamin Harke, and Alberto Diaspro, "Frequency dependent detection in a STED microscope using modulated excitation light," Opt. Express 21, 210-219 (2013)

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