This paper investigated the performance of affine optical flow (AFOF) in motion tracking of the arterial wall from B-mode ultrasound images and the effect of its combination with multiscale image analysis on the accuracy of the process. Multiscale AFOF (MAFOF) exploits the information obtained with AFOF from the approximation sub-images at different spatial resolution levels of the images, obtained using a 2D discrete wavelet transform. Both AFOF and MAFOF were evaluated through their application to synthetic image sequences of the common carotid artery. Multiscale image analysis increased the accuracy in motion tracking, with MAFOF yielding average displacement error reductions of 9% with respect to AFOF. The methods were also effectively applied to real ultrasound image sequences of the carotid artery. The results showed that MAFOF could be considered as a reliable estimator for arterial wall motion from B-mode ultrasound images.