The digital cushion (DC) plays a role in absorbing and dampening forces applied to the foot and therefore supports internal structures such as navicular bone; yet, its architecture is not well-known. The goal of this study was to characterize the microanatomical structure of the DC in horses with clinically sound hooves. Both forefeet from the cadavers of 12 adult Quarter horses were cut and sectioned and samples of the following 4 regions of the DC were obtained: axial proximal (AxProx), axial distal (AxDis), abaxial lateral (AbxLat), and abaxial medial (AbxMed). The samples were processed and stained with hematoxylin and eosin, Masson's trichrome, and Weigert's elastic stains. On each slide, 2 central 3- × 3-mm areas were microscopically assessed and all measurements were done within the 9-mm2 area. The number of detected collagen bundles, nerve fascicles, vessels, and the diameter of wall thickness and lumen of blood vessels were measured. Elastic fiber profiles were categorized based on relative density of elastic fibers detected in the field. The percentage of samples in which chondrocytes and adipose tissues were either present or absent was calculated. Significant structural differences were identified among the 4 regions of the DC. The AxDis region contained more collagen bundles (P < 0.0001) and less elastic fiber profiles than the AxProx region (P < 0.0001). The AxDis also contained more collagen bundles than the AbxMed and AbxLat (P < 0.0001) regions. Our findings provide insight into the structure of the DC of mature Quarter horses. The structural differences in the various regions of the DC are presumably related to the different functional properties of those regions; yet more research is warranted.