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Journal of the Optical Society of America A

Journal of the Optical Society of America A

| OPTICS, IMAGE SCIENCE, AND VISION

  • Editor: Steven A. Burns
  • Vol. 24, Iss. 9 — Sep. 1, 2007
  • pp: 2622–2627

Enhanced three-dimensional deconvolution microscopy using a measured depth-varying point-spread function

Joshua W. Shaevitz and Daniel A. Fletcher  »View Author Affiliations


JOSA A, Vol. 24, Issue 9, pp. 2622-2627 (2007)
http://dx.doi.org/10.1364/JOSAA.24.002622


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Abstract

We present a technique to systematically measure the change in the blurring function of an optical microscope with distance between the source and the coverglass (the depth) and demonstrate its utility in three-dimensional (3D) deconvolution. By controlling the axial positions of the microscope stage and an optically trapped bead independently, we can record the 3D blurring function at different depths. We find that the peak intensity collected from a single bead decreases with depth and that the width of the axial, but not the lateral, profile increases with depth. We present simple convolution and deconvolution algorithms that use the full depth-varying point-spread functions and use these to demonstrate a reduction of elongation artifacts in a reconstructed image of a 2 μ m sphere.

© 2007 Optical Society of America

OCIS Codes
(100.3020) Image processing : Image reconstruction-restoration
(100.6890) Image processing : Three-dimensional image processing
(110.0180) Imaging systems : Microscopy
(140.7010) Lasers and laser optics : Laser trapping
(170.6900) Medical optics and biotechnology : Three-dimensional microscopy
(180.2520) Microscopy : Fluorescence microscopy

ToC Category:
Image Processing

History
Original Manuscript: February 26, 2007
Revised Manuscript: April 23, 2007
Manuscript Accepted: May 1, 2007
Published: July 27, 2007

Virtual Issues
Vol. 2, Iss. 10 Virtual Journal for Biomedical Optics

Citation
Joshua W. Shaevitz and Daniel A. Fletcher, "Enhanced three-dimensional deconvolution microscopy using a measured depth-varying point-spread function," J. Opt. Soc. Am. A 24, 2622-2627 (2007)
http://www.opticsinfobase.org/josaa/abstract.cfm?URI=josaa-24-9-2622


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