A novel digital radiographic method has been developed for measuring the thickness of a tissue sample by iodine displacement. This simple, accurate method is useful both in medical research and in the comparison of pathological and clinical findings. Radiographic measurements of tissue samples in air suffer from the limitation that the quantity measured depends on the product of both thickness and radiographic attenuation coefficient. This technique allows one to obtain thickness measurements from a digital radiograph of a tissue sample suspended in a bath of radio-opaque contrast agent. The attenuation of the iodinated contrast agent is much higher than that of tissue or calcium. Thus the resulting image is determined largely by the contour of the surrounding bath, and is only slightly influenced by the composition of the tissue. This technique improves both the accuracy and precision of radiographic thickness determination. In this paper the iodine displacement technique is described and the accuracy and precision of thickness measurements in appropriate phantoms are quantified. This technique has been used with both image intensifier and screen-film based imaging systems to obtain thickness maps of calcified human aorta, with precision better than 4% and spatial resolution of 2.5 mm-1.