Estimation of 3D point spread function of a widefield microscope by stochastic least square minimization method

S. S. Goutam Buddha; Nagendra Kumar; Santanu Konwar; Bosanta R. Boruah

doi:10.1117/12.2651025

16 May 2023 Estimation of 3D point spread function of a widefield microscope by stochastic least square minimization method

S. S. Goutam Buddha, Nagendra Kumar, Santanu Konwar, Bosanta R. Boruah

Author Affiliations +

Proceedings Volume 12385, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXX; 123850C (2023) https://doi.org/10.1117/12.2651025
Event: SPIE BiOS, 2023, San Francisco, California, United States

Conference Poster

Abstract

The point-spread function (PSF) plays a fundamental role in deciding the resolution of an optical microscope. One way to find the experimental PSF is to image sub-resolution nanobead. However, the PSF can also be approximated numerically by knowing the optical arrangement of the imaging system. In this paper, we estimate the experimental three-dimensional PSF of a widefield microscope by taking several z-stacks of an arbitrary target, followed by application of our 2D PSF estimation scheme to each z slice. Here we use a standard resolution test target as the arbitrary target whose geometrical-optics predicted image can be easily constructed.

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S. S. Goutam Buddha, Nagendra Kumar, Santanu Konwar, and Bosanta R. Boruah "Estimation of 3D point spread function of a widefield microscope by stochastic least square minimization method", Proc. SPIE 12385, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXX, 123850C (16 May 2023); https://doi.org/10.1117/12.2651025

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