(a) Clinical and (b) H&E images of the lesion. (c) Cross-sectional OCT image showing a raised lesion with increased density of blood vessels. Bar = 1 mm. (d) Enface (horizontal) RCM mosaic (mm-scale) image showing the overall morphology of the lesion at a depth of 35 μm (spinosum layer). Bar = 500 μm. (e) 3D RCM images enclosing the area outlined in the RCM image in d, revealing migrating melanocytes in the spinosum layer (arrowheads, z = 25 μm, 35 μm), epidermal disarray (z = 25–75 μm), and nests of melanoma cells at the DEJ (arrowheads, z = 75 μm). Bar = 100 μm. (f) 3D MPM images acquired at the same location showing migrating melanocytes in the spinosum layer (arrowheads, z = 25 μm, 35 μm), epidermal disarray (z = 25–75 μm), and a nest of melanoma cells (arrowheads) surrounded by collagen fibers (blue) at the DEJ (z = 75 μm). Bar = 40 μm. 3D, three-dimensional; DEJ, dermal–epidermal junction; MPM, multiphoton microscopy; OCT, optical coherence tomography; RCM, reflectance confocal microscopy.