1Tata Institute of Fundamental Research (India) 2Stanford Univ. (United States) 3Abhayapuri College (India) 4Indian Institute of Technology Guwahati (India)
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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, 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