The present invention provides methods, a system, and a program product for the rapid detection and correction of spherical aberration in microscopy systems. More specifically, the present invention empirically derives a pupil function, adaptively corrects PSF parameters, and automatically detects the coefficient for spherical aberration. A first aspect of the invention provides a method for detecting and estimating spherical aberration in an acquired image obtained using an optical system, comprising the steps of deconvolving an image using each of a plurality of point spread functions, wherein each point spread function has a different spherical aberration value, calculating an image energy for each deconvolved image, and choosing as a spherical aberration coefficient the spherical aberration value corresponding to the deconvolved image having the lowest image energy, wherein a spherical aberration coefficient other than 0 indicates the presence of spherical aberration in the acquired image and its distance from 0 is an estimation of the degree and direction of spherical aberration.