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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 15, Iss. 6 — Mar. 19, 2007
  • pp: 3361–3371

Efficient fluorescence inhibition patterns for RESOLFT microscopy

Jan Keller, Andreas Schönle, and Stefan W. Hell  »View Author Affiliations


Optics Express, Vol. 15, Issue 6, pp. 3361-3371 (2007)
http://dx.doi.org/10.1364/OE.15.003361


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Abstract

By exploiting the saturation of a reversible single photon transition, RESOLFT microscopy is capable of resolving three dimensional structures inside specimen with a resolution that is no longer limited by the wavelength of the light in use. The transition is driven by a spatially varying intensity distribution that features at least one isolated point, line or plane with zero intensity and the resolution achieved depends critically on the field distribution around these zeros. Based on a vectorial analysis of the image formation in a RESOLFT microscope, we develop a method to effectively search for optimal zero intensity point patterns under typical experimental conditions. Using this approach, we derived a spatial intensity distribution that optimizes the focal plane resolution. Moreover, we outline a general strategy that allows optimization of the resolution for a given experimental situation and present solutions for the most common cases in biological imaging.

© 2007 Optical Society of America

OCIS Codes
(110.0180) Imaging systems : Microscopy
(110.2990) Imaging systems : Image formation theory
(180.2520) Microscopy : Fluorescence microscopy
(220.1230) Optical design and fabrication : Apodization

ToC Category:
Microscopy

History
Original Manuscript: January 10, 2007
Revised Manuscript: February 16, 2007
Manuscript Accepted: February 26, 2007
Published: March 19, 2007

Virtual Issues
Vol. 2, Iss. 4 Virtual Journal for Biomedical Optics

Citation
Jan Keller, Andreas Schönle, and Stefan W. Hell, "Efficient fluorescence inhibition patterns for RESOLFT microscopy," Opt. Express 15, 3361-3371 (2007)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-15-6-3361


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