Pulse diagnosis is an important technique in traditional Chinese (veterinary) medicine, wherein acupuncture (AP) treatment is adjusted according to pulse changes (blood flow). This diagnosis, however, can be subjective. Currently no quantitative evaluation method of blood flow secondary to acupoint stimulation has been established. The objective of this research was to apply ultrasound color Doppler imaging (UCDI) to assess the effect of AP at LU-9 and PC-6 on blood flow. Thirty-two healthy dogs were enrolled, each receiving five different AP treatments (≥1 week apart): AP at LU-9; AP at PC-6; AP at LU-9 and PC-6; AP at a sham point; and no AP (waiting in room). Before and after each treatment, the same investigator, blinded to AP treatment, performed UCDI to record blood flow data. Outcome data included vessel diameter (VD), peak systolic velocity (PSV), end-diastolic velocity (EDV), time average maximum velocity (TAMAX), and pulsatility indices (PI). Additionally, blood flow calculated endpoints included: resistive index (RI), cross-sectional area (CSA), and blood flow volume (BFV). The data showed that AP at PC-6 had a statistically significant impact on blood flow parameters: VD (p=0.002), PSV (p=0.001), EDV (p=0.005), TAMAX (p<0.001), CSA (p=0.005), and BFV (p<0.001). The findings from this study suggest that AP treatment could increase blood flow, especially with respect to flow velocity and volume, dependent on acupoint used. Clinical application of the Doppler ultrasound technique used in this study, allows monitoring of acupuncture-associated blood flow changes, leading to development of unique therapeutic approaches to manage canine vascular and cardiac diseases.