A substrate has three or more overlay gratings formed thereon by a lithographic process. Each overlay grating has a known overlay bias. The values of overlay bias include for example two values in a region centered on zero and two values in a region centered on P/2, where P is the pitch of the gratings. Overlay is calculated from asymmetry measurements for the gratings using knowledge of the different overlay bias values and an assumed non-linear relationship between overlay and target asymmetry, thereby to correct for feature asymmetry. The periodic relationship in the region of zero bias and P/2 has gradients of opposite sign. The calculation allows said gradients to have different magnitudes as well as opposite sign. The calculation also provides information on feature asymmetry and other processing effects. This information is used to improve subsequent performance of the measurement process, and/or the lithographic process.