A study of Gaussian approximations of fluorescence microscopy PSF models

Bo Zhang; Josiane Zerubia; Jean-Christophe Olivo-Marin

doi:10.1117/12.645650

23 February 2006 A study of Gaussian approximations of fluorescence microscopy PSF models

Bo Zhang, Josiane Zerubia, Jean-Christophe Olivo-Marin

Proceedings Volume 6090, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII; 60900K (2006) https://doi.org/10.1117/12.645650
Event: SPIE BiOS, 2006, San Jose, California, United States

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%).

Citation Download Citation

Bo Zhang, Josiane Zerubia, and Jean-Christophe Olivo-Marin "A study of Gaussian approximations of fluorescence microscopy PSF models", Proc. SPIE 6090, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII, 60900K (23 February 2006); https://doi.org/10.1117/12.645650

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available