Volume scanning electron microscopy (VSEM) involves the serial sectioning and imaging of a sample using scanning electron microscopy (SEM), followed by segmentation and three-dimensional (3D) reconstruction using computer software packages to allow visualization of 3D structures. VSEM can reveal qualitative and quantitative properties of organelles and cells within tissues at nanoscale. The ability to visualize spatial relationships of structures of interest within and across cells in 3D space in particular sets VSEM apart from conventional SEM and transmission electron microscopy. Here, we provide an overview of VSEM platforms and image processing, highlighting characteristics that will aid selection of a method to address specific research questions in dermatological research.