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Proceedings Paper

A study of Gaussian approximations of fluorescence microscopy PSF models
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Paper Abstract

Despite the availability of rigorous physical models of microscopy point spread functions (PSFs), approximative PSFs, particularly separable Gaussian approximations are widely used in practical microscopic data processing. In fact, compared with a physical PSF model, which usually involves non-trivial terms such as integrals and infinite series, a Gaussian function has the advantage that it is much simpler and can be computed much faster. Moreover, due to its special analytical form, a Gaussian PSF is often preferred to facilitate the analysis of theoretical models such as Fluorescence Recovery After Photobleaching (FRAP) process and of processing algorithms such as EM deconvolution. However, in these works, the selection of Gaussian parameters and the approximation accuracy were rarely investigated. In this paper, we present a comprehensive study of Gaussian approximations for diffraction-limited 2D/3D paraxial/non-paraxial PSFs of Wide Field Fluorescence Microscopy (WFFM), Laser Scanning Confocal Microscopy (LSCM) and Disk Scanning Confocal Microscopy (DSCM) described using the Debye integral. Besides providing an optimal Gaussian parameter for the 2D paraxial WFFM PSF case, we further derive nearly optimal parameters in explicit forms for each of the other cases, based on Maclaurin series matching. Numerical results show that the accuracy of the 2D approximations is very high (Relative Squared Error (RSE) < 2% in WFFM, < 0.3% in LSCM and < 4% in DSCM). For the 3D PSFs, the approximations are average in WFFM (RSE ≃ 16-20%), accurate in DSCM (RSE≃ 3-6%) and nearly perfect in LSCM (RSE ≃ 0.3-0.5%).

Paper Details

Date Published: 23 February 2006
PDF: 11 pages
Proc. SPIE 6090, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII, 60900K (23 February 2006); doi: 10.1117/12.645650
Show Author Affiliations
Bo Zhang, Institut Pasteur (France)
École Nationale Supérieure des Télécommunications (France)
Josiane Zerubia, Institut National de Recherche en Informatique et en Automatique (France)
Jean-Christophe Olivo-Marin, Institut Pasteur (France)

Published in SPIE Proceedings Vol. 6090:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson, Editor(s)

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