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Proceedings Paper

Application of phase-retrieved pupil functions in wide-field fluorescence microscopy
Author(s): Bridget M. Hanser; Mats G. L. Gustafsson; David A. Agard; John W. Sedat
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Paper Abstract

Multidimensional fluorescence microscopy data are often collected under imaging conditions that cause aberrations. For proper deconvolution of such data, one needs to generate a Point Spread Function (PSF) that includes the aberrations. Even if it is possible to model the imaging aberrations theoretically, there remains the problem of modifying the PSF accordingly. Measured PSFs are difficult to modify, and theoretical PSFs lack lens- and system-specific features. Phase retrieval approaches can be used to produce a compact and modifiable description of a microscope system, including its measured, system-specific performance. These pupil functions can be used in a depth-dependent deconvolution to correct for geometric distortions and improve restoration of relative intensities compared to spatially invariant deconvolution using unaberrated PSFs.

Paper Details

Date Published: 15 May 2002
PDF: 7 pages
Proc. SPIE 4621, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX, (15 May 2002); doi: 10.1117/12.467844
Show Author Affiliations
Bridget M. Hanser, Univ. of California/San Francisco (United States)
Mats G. L. Gustafsson, Univ. of California/San Francisco (United States)
David A. Agard, Univ. of California/San Francisco (United States)
John W. Sedat, Univ. of California/San Francisco (United States)


Published in SPIE Proceedings Vol. 4621:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson, Editor(s)

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