Intima-media thickness (IMT) measurements have gained increasing attention, because IMT is assumed to represent the endothelial adaptive response to physiological and pathophysiological processes. The main aim of the present study was to assess the intrasubject intrasession variability of a new off-line automated radio frequency (RF) IMT method in comparison with an already established off-line manual B-mode IMT method. IMT also was assessed by means of an on-line manual B-mode and an on-line manual RF IMT method. We investigated posterior wall IMT 0-1 cm proximal to the bulb in both common carotid arteries of 16 young (20-31 y; mean 25 y) female and male and 13 elderly (51-65 y; mean 56 y) female volunteers. Two commercially available ultrasound devices (Pie Medical Scanner 200 and Ultramark 9) were used to assess the effects of signal processing on the off-line automated RF IMT method. Intrasubject intrasession variability was determined using the standard deviation to evaluate and compare the various methods. Spearman rank correlation coefficients and Bland and Altman bias and limits of agreement were calculated to objectivate the comparability between the various methods. Intrasubject intrasession variation of IMT estimates was not statistically different between any of the methods. We observed a good comparability between the commonly used off-line manual B-mode IMT method and the off-line automated RF IMT method at the level of the common carotid artery.