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In this paper we propose a point spread function (PSF) estimation scheme for an optical microscope. The scheme is based on minimizing the least square error between the image of an arbitrary target and the convolution of a stochastically constructed PSF array with the geometrical-optics predicted image of the target. The proposed scheme is independent of the geometry of the imaging system and will work in any type of optical microscopes. The scheme enables PSF estimation without requiring any modification of the microscope setup. We have demonstrated the proposed scheme both numerically and experimentally.

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