Although the gross mechanical properties of ageing tissues have been extensively documented, biological tissues are highly heterogeneous and little is known concerning the variation of micro-mechanical properties within tissues. Here, we use Scanning Acoustic Microscopy (SAM) to map the acoustic wave speed (a measure of stiffness) as a function of distance from the outer adventitial layer of cryo-sectioned ferret aorta. With a 400 MHz lens, the images of the aorta samples matched those obtained following chemical fixation and staining of sections which were viewed with fluorescence microscopy. Quantitative analysis was conducted with a frequency scanning or V(f) technique by imaging the tissue from 960 MHz to 1.1 GHz. Undulating acoustic wave speed (stiffness) distributions corresponded with elastic fibre locations in the tissue; there was a decrease in wave speed of around 40 ms-1 from the adventitia (outer layer) to the intima (innermost).