Overlay error of a lithographic process is measured using a plurality of target structures, each target structure having a known overlay bias. A detection system captures a plurality of images (740) representing selected portions of radiation diffracted by the target structures under a plurality of different capture conditions (λ1, λ2). Pixel values of the captured images are combined (748) to obtain one or more synthesized images (750). A plurality of synthesized diffraction signals are extracted (744) from the synthesized image or images, and used to calculate a measurement of overlay. The computational burden is reduced compared with extracting diffraction signals from the captured images individually. The captured images may be dark-field images or pupil images, obtained using a scatterometer.