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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 20, Iss. 22 — Oct. 22, 2012
  • pp: 24585–24599

Flexible structured illumination microscope with a programmable illumination array

Pavel Křížek, Ivan Raška, and Guy M. Hagen  »View Author Affiliations


Optics Express, Vol. 20, Issue 22, pp. 24585-24599 (2012)
http://dx.doi.org/10.1364/OE.20.024585


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Abstract

Structured illumination microscopy (SIM) has grown into a family of methods which achieve optical sectioning, resolution beyond the Abbe limit, or a combination of both effects in optical microscopy. SIM techniques rely on illumination of a sample with patterns of light which must be shifted between each acquired image. The patterns are typically created with physical gratings or masks, and the final optically sectioned or high resolution image is obtained computationally after data acquisition. We used a flexible, high speed ferroelectric liquid crystal microdisplay for definition of the illumination pattern coupled with widefield detection. Focusing on optical sectioning, we developed a unique and highly accurate calibration approach which allowed us to determine a mathematical model describing the mapping of the illumination pattern from the microdisplay to the camera sensor. This is important for higher performance image processing methods such as scaled subtraction of the out of focus light, which require knowledge of the illumination pattern position in the acquired data. We evaluated the signal to noise ratio and the sectioning ability of the reconstructed images for several data processing methods and illumination patterns with a wide range of spatial frequencies. We present our results on a thin fluorescent layer sample and also on biological samples, where we achieved thinner optical sections than either confocal laser scanning or spinning disk microscopes.

© 2012 OSA

OCIS Codes
(100.3010) Image processing : Image reconstruction techniques
(180.1790) Microscopy : Confocal microscopy
(180.2520) Microscopy : Fluorescence microscopy
(230.6120) Optical devices : Spatial light modulators
(150.1488) Machine vision : Calibration

ToC Category:
Microscopy

History
Original Manuscript: August 27, 2012
Revised Manuscript: September 26, 2012
Manuscript Accepted: September 28, 2012
Published: October 12, 2012

Citation
Pavel Křížek, Ivan Raška, and Guy M. Hagen, "Flexible structured illumination microscope with a programmable illumination array," Opt. Express 20, 24585-24599 (2012)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-20-22-24585


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