This study aimed to morphologically evaluate the stratum corneum (SC) barrier function. Acute SC barrier dysfunction models were produced by tape stripping (TS) in five dogs. Using TS keratinocytes, the size of cells, multilayered ratio, nuclear cell ratio, and sulfhydryl group ratio were measured together with transepidermal water loss (TEWL). TEWL was significantly increased after TS injury compared to before injury, and the significantly increased level remained at 7 days post-injury. The TEWL values at 14 and 28 days post-injury were the same as before injury. The size of keratinocytes significantly decreased after TS injury. However, no significant difference compared to before injury was found at 7, 14, or 28 days post-injury. The multilayered ratio was significantly elevated after TS injury compared to before injury, and this significantly elevated level was maintained at 7 days post-injury. However, the levels at 14 and 28 days post-injury were the same as before injury. The nuclear cell and sulfhydryl group ratios were significantly higher after TS injury, and remained significantly elevated at 7 and 14 days post-injury. However, they showed no significant difference from before TS injury at 28 days post-injury. In the present study, the measured indicators demonstrated a significant correlation with TEWL. The correlation coefficients were as follows: r=–0.651, cell size; r=0.809, the multilayered ratio; r=0.464, the nuclear cell ratio; and r=0.726, the sulfhydryl group ratio. Morphological assessment via TS keratinocytes is useful for evaluating the SC barrier function in dogs.