Optimal determination of an overlay target using machine learning

There are provided systems and methods comprising, for each given overlay target of a plurality of different overlay targets to be manufactured on a semiconductor specimen, said given overlay target comprising a plurality of stacked semiconductor layers, obtaining a design image of the given overlay target, feeding the design image to a trained machine learning model, to simulate at least one image of the given overlay target that would have been acquired by an electron beam examination system, using the at least one image to determine, before actual manufacturing of the given overlay target, data informative of at least one simulated overlay in the image, and using the data informative of the at least one simulated overlay of each given overlay target to select at least one optimal overlay target among the plurality of different overlay targets, the optimal overlay target being usable to be manufactured on the semiconductor specimen.