Atopic dermatitis is a dermatological disease in which dogs are genetically predisposed to skin inflammation and allergies, with pruritus being the first clinical sign to be manifested, with itchy and eczematoid lesions. It is believed that affected dogs present a deficient cutaneous barrier and ceramides loss and that the loss of ceramides would be the cause of increased allergens adsorption and the trigger for immunological reactions and exaggerated inflammation. This study aimed to compare the quantity of epidermal lipids in normal and atopic dogs before and after treatment with ceramides solution. Ten dogs attended at UNESP Veterinary Hospital (Botucatu - Brazil) were selected according to the criteria described by Favrot et al. (2010) and submitted to skin histopathological evaluation. Samples were obtained from axillary and cervical areas. After this procedure, animals were treated with topic ceramides solution for 30 days and were again submitted for histological skin evaluation. All samples were processed by frozen section at -25°C and 20 µm and stained by Oil Red O and Sudan III, previously standardized for lipid evaluation in corneal layer. Epidermal width and area were measured before and 30 days after treatment with ceramides by color deconvolution method with ImageJ® freeware. It was possible to observe that atopic animals have significantly thinner lipid epidermal layer than normal dogs and that the lipid layer of atopic animals increased after treatment. After statistical evaluation, it was possible to conclude that the lipid barrier was indeed restored after treatment and that clinical improvement was achieved.