Objective To explore the application value of ultra-high frequency ultrasound in detection of femoral artery stenosis in rabbits. Methods Twenty-four healthy male New Zealand white rabbits (body weight 2.5-3.0 kg) were randomly divided into three groups, 8 rabbits in each group. Preparation of femoral atherosclerosis model: the rabbits were fed with high fat diet for 4 weeks, 8 weeks and 12 weeks after femoral artery balloon injury, respectively. The changes of cholesterol level were observed. We used ultra-high frequency ultrasonic probe to observe the femoral artery, to assess the diameter stenosis rate and peak systolic velocity (SPV) at the symptomatic side, and the histological areal stenosis was evaluated. Results 1. The blood cholesterol levels were increased after fed high cholesterol diet, with a significant difference among the groups (P<0.001); 2. There was a stenosis to a different degree in the modeling artery after fed high cholesterol diet for 4 wks, 8 wks, and 12 wks, and the stenosis degree was increased gradually. At 12 w, the degree of stenosis was higher than that at 4 w and 8 w, respectively, showing statistical significance (P<0.001). At the symptomatic side, the peak systolic flow velocity was increased, and it was higher at 12 w than that at 4w and 8 w, respectively. 3. With the extension of time, the arterial stenosis rate was increased along with the time course (P<0.05). Conclusions 1. High fat diet combined with balloon injury can cause varying degrees of rabbit femoral artery stenosis; 2. Ultrahigh frequency ultrasound can clearly show different degrees of rabbit femoral artery stenosis, and can be used for detecting the lesions in animal models of peripheral vascular diseases.