Rigid registration of a series of ten images collected at −1 µm defocus. The resulting image is blurred as not all beam-induced movements could be accounted for. Panel (b) is an enlargement of the area marked by the red box in panel (a). Panels (c) and (d) [enlargement of (c)] show the non-rigid registration of the same series. The yellow contour in panel (c) shows the region of the image that was corrected. In contrast to (a) and (b), gold particles have become sharp and more protein particles become visible in (c) and (d). The scale bar shown in panels (b) and (d) corresponds to 50 nm.