This study determined the choroidal thickness of senior (SN, n = 24) and middle-aged (MA, n = 17) healthy, mixed-breed mesocephalic dogs, both males (M) and females (F), using spectral domain optical coherence tomography (SD-OCT). The dogs were divided into two groups for examination: MA dogs (4–7 years old; 6 M, 11 F) and SN dogs (8–13 years old; 12 M, 12 F). Choroidal thickness of the dogs was investigated using SD-OCT radial and linear scans. The software of the device allowed determination of the exact measurement location on the choroid. Measurements of the choroid were taken manually using the SD-OCT calliper function at distances of 5,000–6,000 μm (dorsal and ventral) and 4,000–7,000 μm (nasal and temporal) from the optic disc. Mean (µm ± SD) (MA, SN) dorsal (188 ± 28, 184 ± 33), ventral (116 ± 23, 111 ± 16), temporal (152 ± 31, 151 ± 26), and nasal (135 ± 27, 132 ± 18) choroidal thicknesses demonstrated significant differences (P < 0.02–0.001) between all areas within each group. The choroid was thickest in the dorsal region and thinnest in the ventral region. There were no significant differences based on age. Mean (µm ± SD) (M, F) dorsal (181 ± 32, 190 ± 30), ventral (117 ± 16, 11 ± 21), temporal (150 ± 26, 153 ± 30), and nasal (128 ± 20, 138 ± 23) choroidal thicknesses demonstrated significant differences (P < 0.05) between dorsal and nasal regions. The choroidal thickness in dogs depends on the area assessed independently of their age and sex.