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Proceedings Paper

Quantitative depth-variant imaging for fluorescence microscopy using the COSMOS software package
Author(s): Chrysanthe Preza; Vimeetha Myneni
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Paper Abstract

The previously developed depth-variant expectation maximization (DVEM) restoration algorithm for fluorescent microscopy [1] has been implemented as part of the computational optical sectioning microscopy open source (COSMOS) software package [2]. COSMOS facilitates the development and dissemination of the DVEM algorithm which addresses depth-variant imaging due to spherical aberrations. Physical parameters such as the imaging lens, the size of the object and its refractive index contribute to the amount of spherical aberrations present in the image. The algorithm is based on a depth-variant imaging model and it requires a small number of point-spread functions (PSFs) defined at different depths within the sample. An acceptable choice for the number of PSFs needed for the computation provides a tradeoff between accuracy of the result and computational complexity. In this paper we show results obtained from simulation studies in which the performance of the DVEM algorithm is investigated for different parameters that contribute to depth variability. Some of the DVEM algorithm results are compared to results obtained with a spaceinvariant (or deconvolution) method from the same simulated data. The main conclusion from these studies is that a small number of PSFs can be used for the estimation without a significant loss in algorithm performance rendering the algorithm suitable for practical use. Furthermore, DVEM offers improvements over deconvolution methods with a small increase in computational complexity.

Paper Details

Date Published: 24 February 2010
PDF: 8 pages
Proc. SPIE 7570, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVII, 757003 (24 February 2010); doi: 10.1117/12.840688
Show Author Affiliations
Chrysanthe Preza, The Univ. of Memphis (United States)
Vimeetha Myneni, The Univ. of Memphis (United States)


Published in SPIE Proceedings Vol. 7570:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVII
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson; Thomas G. Brown, Editor(s)

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