An optimized model of point spread function of microscopy based on fluorescence beads

LU Weimin; YANG Xibin; WEN Gang; ZHENG Xianliang; LI Hui; XIONG Daxi

doi:10.7510/jgjs.issn.1001-3806.2016.05.005

In order to eliminate effect of fluorescent bead diameter on measurement results in traditional microscopic point spread function measurement method and improve the measurement accuracy, 2-D equivalent concentration distribution of fluorescent beads was deduced based on theoretical simulation and least square fitting method. The imaging process of fluorescent beads was simulated. The relationships among fluorescent beads diameter, full width at half maximum (FWHM) of fluorescent beads intensity distribution and FWHM of system point spread function were analyzed by using the least square fitting method and residual error fitting method. Precise FWHM correction model of point spread function was obtained. The results show that the relative error of point spread function FWHM is about 1% when measuring system point spread function with 100nm fluorescent beads. The precise measurement of point spread function is realizable with the optimized model.

An optimized model of point spread function of microscopy based on fluorescence beads

Corresponding author: XIONG Daxi, xiongdx@sibet.ac.cn

1. Jiangsu Key Laboratory of Medical Optics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, China;

2. Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun 130033, China;

3. College of Materials Science and Opto-Electronic Technology, University of Chinese Academy of Sciences, Beijing 100039, China

Received Date: 2015-07-07

Accepted Date: 2015-07-16

Available Online: 2016-09-25

Abstract: In order to eliminate effect of fluorescent bead diameter on measurement results in traditional microscopic point spread function measurement method and improve the measurement accuracy, 2-D equivalent concentration distribution of fluorescent beads was deduced based on theoretical simulation and least square fitting method. The imaging process of fluorescent beads was simulated. The relationships among fluorescent beads diameter, full width at half maximum (FWHM) of fluorescent beads intensity distribution and FWHM of system point spread function were analyzed by using the least square fitting method and residual error fitting method. Precise FWHM correction model of point spread function was obtained. The results show that the relative error of point spread function FWHM is about 1% when measuring system point spread function with 100nm fluorescent beads. The precise measurement of point spread function is realizable with the optimized model.

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Reference (17)

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An optimized model of point spread function of microscopy based on fluorescence beads

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