Abstract: In a method of determining the focus of a lithographic apparatus used in a lithographic process on a substrate, the lithographic process is used to form a structure on the substrate, the structure having at least one feature which has an asymmetry in the printed profile which varies as a function of the focus of the lithographic apparatus on the substrate. A first image of the periodic structure is formed and detected while illuminating the structure with a first beam of radiation. The first image is formed using a first part of non-zero order diffracted radiation. A second image of the periodic structure is foamed and detected while illuminating the structure with a second beam of radiation. The second image is formed using a second part of the non-zero order diffracted radiation which is symmetrically opposite to the first part in a diffraction spectrum.