The determination of the modulus of the pupil function in reflection acoustic and optical microscopy with the help of spherical particles is demonstrated. The theoretical examination shows that this method can give good results if the pupil function drops smoothly towards the edge. For a pupil with a sharp edge this technique can give the accurate pupil function only by imaging large spheres. The dependence of the accuracy of the method on the size of the spherical particle is analyzed. Numerical and experimental results obtained for the confocal scanning optical microscope and the scanning acoustic microscope are examined.