We set out to determine microscopic characteristics of the Descemet membrane interface during Descemet membrane endothelial keratoplasty (DMEK) graft preparation. Ten corneas were partially prepared, preserving half of the Descemet membrane attached to the stroma to enable visualisation of the Descemet-stroma interface. This tissue was prepared for viewing with a scanning electron microscope. The Descemet-stroma interface was categorised into three regions: centre, mid-periphery and periphery. We classified adhesions in these regions as either minor thread-like adhesions or major bridge-like adhesions with stromal detachments. We found a region-specific differentiation of the Descemet-stroma morphology. The presence of minor (P = 0,0001) and major (P = 0,0001) adhesions at the explored regions of the Descemet-stroma interface were found to be statistically significant. Fibrotic linear adhesions were predominant in the centre and mid-periphery, whereas the larger bridge-like adhesions were found mainly in the periphery. In addition, we observed a positive correlation between the size of the adhesions and the presence of ruptures in the underlying stromal bed. Viewing of the Descemet-stroma interface with electron microscopy reveals morphological differences between the centre of a graft and its periphery. These findings are of potential clinical relevance in terms of developing a better understanding of tissue behaviour during graft preparation.