Gaussian approximations of fluorescence microscope point-spread function models.
We comprehensively study the least-squares Gaussian approximations of the diffraction-limited 2D-3D paraxial-nonparaxial point-spread functions (PSFs) of the wide field fluorescence microscope (WFFM), the laser scanning confocal microscope (LSCM), and the disk scanning confocal microscope (DSCM). The PSFs are expressed using the Debye integral. Under an L(infinity) constraint imposing peak matching, optimal and near-optimal Gaussian parameters are derived for the PSFs. With an L1 constraint imposing energy conservation, an optimal Gaussian parameter is derived for the 2D paraxial WFFM PSF. We found that (1) the 2D approximations are all very accurate; (2) no accurate Gaussian approximation exists for 3D WFFM PSFs; and (3) with typical pinhole sizes, the 3D approximations are accurate for the DSCM and nearly perfect for the LSCM. All the Gaussian parameters derived in this study are in explicit analytical form, allowing their direct use in practical applications.
Zhang
Bo
B
Unité d'Analyse d'Images Quantitative, Institut Pasteur, Cedex, France. bzhang@pasteur.fr
Zerubia
Josiane
J
Olivo-Marin
Jean-Christophe
JC
D000465:Algorithms; D003198:Computer Simulation; D007089:Image Enhancement / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008954:Models, Biological; D015233:Models, Statistical; D016011:Normal Distribution; D011829:Radiation Dosage; D011874:Radiometry; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity