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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 20, Iss. 7 — Mar. 26, 2012
  • pp: 7362–7374

Strategies to maximize the performance of a STED microscope

Silvia Galiani, Benjamin Harke, Giuseppe Vicidomini, Gabriele Lignani, Fabio Benfenati, Alberto Diaspro, and Paolo Bianchini  »View Author Affiliations


Optics Express, Vol. 20, Issue 7, pp. 7362-7374 (2012)
http://dx.doi.org/10.1364/OE.20.007362


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Abstract

In stimulated emission depletion (STED) microscopy, the spatial resolution scales as the inverse square root of the STED beam's intensity. However, to fully exploit the maximum effective resolution achievable for a given STED beam’s intensity, several experimental precautions have to be considered. We focus our attention on the temporal alignment between the excitation and STED pulses and the polarization state of the STED beam. We present a simple theoretical framework that help to explain their influence on the performance of a STED microscope and we validate the results by imaging calibration and biological samples with a custom made STED architecture based on a supercontinuum laser source. We also highlight the advantages of using time gating detection in terms of temporal alignment.

© 2012 OSA

OCIS Codes
(000.2170) General : Equipment and techniques
(180.2520) Microscopy : Fluorescence microscopy
(350.5730) Other areas of optics : Resolution

ToC Category:
Microscopy

History
Original Manuscript: January 13, 2012
Revised Manuscript: February 26, 2012
Manuscript Accepted: March 1, 2012
Published: March 15, 2012

Virtual Issues
Vol. 7, Iss. 5 Virtual Journal for Biomedical Optics

Citation
Silvia Galiani, Benjamin Harke, Giuseppe Vicidomini, Gabriele Lignani, Fabio Benfenati, Alberto Diaspro, and Paolo Bianchini, "Strategies to maximize the performance of a STED microscope," Opt. Express 20, 7362-7374 (2012)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-20-7-7362


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