A substrate has first and second target structures formed by a lithographic process. Each target structure has a two-dimensional periodic structure formed in a single layer using first and second lithographic steps. The first target structure has features defined in the second lithographic step displaced relative to features defined in the first lithographic step by a first bias amount. The second target structure has features defined in the second lithographic step displaced relative to features defined in the first lithographic step by a second bias amount. An angle-resolved scatter spectrum of the first target structure and an angle-resolved scatter spectrum of the second target structure is obtained. A measurement of a parameter of a lithographic process is derived from the measurements using asymmetry found in the scatter spectra of the first and second target structures.