The accuracy of an algorithm for the automated tracking of tendon excursion from ultrasound images was tested in three experiments. Because the automated method could not be tested against direct measurements of tendon excursion in vivo, an indirect validation procedure was employed. In one experiment, a wire “phantom” was moved a known distance across the ultrasound probe and the automated tracking results were compared with the known distance. The excursion of the musculotendinous junction of the gastrocnemius during frontal and sagittal plane movement of the ankle was assessed in a single cadaver specimen both by manual tracking and with a cable extensometer sutured to the gastrocnemius muscle. A third experiment involved estimation of Achilles tendon excursion in vivo with both manual and automated tracking. Root mean squared (RMS) error was calculated between pairs of measurements after each test. Mean RMS errors of less than 1 mm were observed for the phantom experiments. For the in vitro experiment, mean RMS errors of 8–9% of the total tendon excursion were observed. Mean RMS errors of 6–8% of the total tendon excursion were found in vivo. The results indicate that the proposed algorithm accurately tracks Achilles tendon excursion, but further testing is necessary to determine its general applicability.