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Optics Letters

Optics Letters


  • Editor: Alan E. Willner
  • Vol. 33, Iss. 16 — Aug. 15, 2008
  • pp: 1819–1821

Wide-field fluorescence sectioning with hybrid speckle and uniform-illumination microscopy

Daryl Lim, Kengyeh K. Chu, and Jerome Mertz  »View Author Affiliations

Optics Letters, Vol. 33, Issue 16, pp. 1819-1821 (2008)

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We describe a method of obtaining optical sectioning with a standard wide-field fluorescence microscope. The method involves acquiring two images, one with nonuniform illumination (in our case, speckle) and another with uniform illumination (in our case, randomized speckle). An evaluation of the local contrast in the speckle-illumination image provides an optically sectioned image with low resolution. This is complemented with high-resolution information obtained from the uniform-illumination image. A fusion of both images leads to a full resolution image that is optically sectioned across all spatial frequencies. This hybrid illumination method is fast, robust, and generalizable to a variety of illumination and imaging configurations.

© 2008 Optical Society of America

OCIS Codes
(180.1790) Microscopy : Confocal microscopy
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:

Original Manuscript: May 22, 2008
Manuscript Accepted: June 13, 2008
Published: August 6, 2008

Virtual Issues
Vol. 3, Iss. 10 Virtual Journal for Biomedical Optics

Daryl Lim, Kengyeh K. Chu, and Jerome Mertz, "Wide-field fluorescence sectioning with hybrid speckle and uniform-illumination microscopy," Opt. Lett. 33, 1819-1821 (2008)

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