Mechanical forces during machine milking of dairy cows evoke circulatory impairment of the teat tissue that may affect the teats' defense mechanisms against mastitis pathogens. Ample research describes dimensional changes of different teat traits after machine milking, whereas reports that describe changes in blood circulation of dairy cows' teats are limited. Therefore, the objectives of this study were to (1) describe changes in teat blood circulation that occur after pre-milking teat stimulation and machine milking and (2) study the effect of 2 different milking liners on machine milking-induced changes in teat blood flow. In a randomized trial, Holstein dairy cows were stratified by parity, stage of lactation, and average daily milk yield during the previous week, and allocated to 1 of 2 treatment groups. Treatment consisted of 1 milking observation with either a round or multisided concave milking liner. Teat scans were taken of the left front and the right hind teats using power Doppler ultrasonography. Imaging occurred before pre-milking udder preparation (T1), after completion of pre-milking udder preparation but before milking-unit attachment (T2), and immediately after unit detachment (T3). Perfusion intensity measurements from teat scans were performed with a commercially available software program. Data from 109 cows were analyzed. A general linear mixed model showed differences in perfusion intensity between time points. Least squares means (95% confidence intervals) for T1, T2, and T3, respectively, were 0.035% (0.026-0.047), 0.124% (0.093-0.164), and 0.095% (0.073-0.124). Conversely, no statistically significant differences between treatment groups were observed. We conclude that teat blood circulation is subjected to several influences, including inherent circulatory regulation mechanisms, as well as extrinsic factors such as machine milking. Future research is warranted to decipher the magnitude of their influence and to further our understanding of how these changes relate to the susceptibility to intramammary infection and milking performance.