In an optical inspection tool, an illumination aperture is opened at each of a plurality of aperture positions of an illumination pupil area one at a time across the illumination pupil area. For each aperture opening position, an incident beam is directed towards the illumination pupil area so as to selectively pass a corresponding ray bundle of the illumination beam at a corresponding set of one or more incident angles towards the sample and an output beam, which is emitted from the sample in response to the corresponding ray bundle of the incident beam impinging on the sample at the corresponding set of one or more incident angles, is detected. A defect detection characteristic for each aperture position is determined based on the output beam detected for each aperture position. An optimum aperture configuration is determined based on the determined defect detection characteristic for each aperture position.