(a, b, c) Initial FIB (a), SEM (b) and LM (c) images. (d) Milling of straight edges in the sample allows improved precision of correlation between images acquired in different modalities. Here, the overlay between LM and FIB images of the pre-milled sample is shown. (d, e, f, g) Pairs of SEM and optical microscopy images of the initial sample state (d), two intermediate verification steps (e and f), and the final result (g). Based on the fluorescence signal, 1.5 µm thick lamellae were isolated around the neuronal body (g). Inset in (g) shows a magnified image of a lamella ready for a cryo-lift-out.