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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 15, Iss. 24 — Nov. 26, 2007
  • pp: 16130–16140

Optimization and characterization of a structured illumination microscope

Frédéric Chasles, Benoît Dubertret, and A. Claude Boccara  »View Author Affiliations


Optics Express, Vol. 15, Issue 24, pp. 16130-16140 (2007)
http://dx.doi.org/10.1364/OE.15.016130


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Abstract

Structured illumination microscopy provides a simple and cheap mean to obtain optical sections of a sample. It can be implemented easily on a regular fluorescent microscope and is a scanning free alternative to confocal microscopy. We have analyzed theoretically the performances of the technique in terms of sectioning strength, resolution enhancement along the optical axis, and signal to background as a function of the objective used and the grid’s characteristics (pitch and contrast). We show that under optimized conditions, the axial resolution can be improved by a factor of 1.5 in comparison with an epifluorescence microscope, and that optical cuts with a thickness of less than 400nm can be obtained with a 1.4 numerical aperture objective. We modified the original grid in-step modulation pattern and used a sinusoidal modulation for the grid displacement. Optical sections are computed by combining four images acquired during one modulation period. This algorithm is very stable even for modulations at high frequencies. The speed of the acquisition is thus only limited by the performance of the detector and the signal/background ratio of the sample. Finally, we compared our technique to commercial setups: a confocal microscope, a Spinning Disk Microscope and a Zeiss Apotome.

© 2007 Optical Society of America

OCIS Codes
(170.6900) Medical optics and biotechnology : Three-dimensional microscopy
(180.0180) Microscopy : Microscopy
(180.2520) Microscopy : Fluorescence microscopy

ToC Category:
Microscopy

History
Original Manuscript: April 6, 2007
Revised Manuscript: June 7, 2007
Manuscript Accepted: June 8, 2007
Published: November 20, 2007

Virtual Issues
Vol. 2, Iss. 12 Virtual Journal for Biomedical Optics

Citation
Frédéric Chasles, Benoit Dubertret, and A. Claude Boccara, "Optimization and characterization of a structured illumination microscope," Opt. Express 15, 16130-16140 (2007)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-15-24-16130


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