Purpose: The subcutaneous fat tissue is separated into 2 layers by the subcutaneous fascia: the superficial and deep fat layers. The two fat layers have different structures according to the body regions. The purpose of this study is to evaluate the distribution and pattern of the two fat layers in the human body by computed tomography (CT) and histological analysis according to age, sex, anatomical region, and body mass index (BMI). Methods: This study included 200 males and 200 females who underwent 64-channel dynamic CT in our hospital. The patients were divided into 5 groups according to 10 years of their ages separately in either male or female gender. The thickness of the superficial and deep fat layers was measured in the abdominal, pelvic, and femoral regions, and we analyzed the values. Statistical analyses were performed using SPSS software. The $3{ imes}3$-cm whole fat layers were harvested from the same sites of 3 cadavers for histological examination, and one cadaver was dissected for gross evaluation. Results: The total thickness of subcutaneous fat tissue was greater in females than in males, and the ratio of the superficial fat layer to the whole fat layer was higher in females. The superficial fat layer became thinner with increasing age in males. As BMI increased, the total fat layer became thicker, and the superficial fat layer became thicker than the deep fat layer. On histological examination, the superficial fat layer had small adipose lobules and showed a densely distributed pattern in the abdominal region, whereas in the femoral region, it had large adipose lobules and showed a sparsely distributed pattern. There were no significant differences in the histological findings of the deep fat layer between the 3 body regions. Conclusion: Significant differences in histological findings of the two fat layers were found in relation to age, sex, anatomical region, and BMI. The superficial fat layer became thinner with increasing age in males, but it was constant in females. As BMI increased, the total thickness of subcutaneous fat tissue became greater, and the superficial fat layer became thicker than the deep fat layer. Our measurements can be used to understand the characteristics of the fat layers in relation to age, sex, anatomical region, and BMI.