On-the-fly estimation of a microscopy point spread function

Estimated vs Measured PSF


A proper estimation of realistic point-spread function (PSF) in optical microscopy can significantly improve the deconvolution performance and assist the microscope calibration process. In this work, by exemplifying 3D wide-field fluorescence microscopy, we propose an approach for estimating the spherically aberrated PSF of a microscope, directly from the observed samples. The PSF, expressed as a linear combination of 4 basis functions, is obtained directly from the acquired image by minimizing a novel criterion, which is derived from the noise statistics in the microscope. We demonstrate the effectiveness of the PSF approximation model and of our estimation method using both simulations and real experiments that were carried out on quantum dots. The principle of our PSF estimation approach is sufficiently flexible to be generalized non-spherical aberrations and other microscope modalities.

Opt. Express, 26(20), 26120-26133, 2018