Purpose: The aim of this study was to evaluate the influence of depth and underlying bone on strain ratios and shear wave speeds for three different muscles in healthy volunteers. For strain ratios the influence from different reference region-of-interest positions was also evaluated. Material and methods: Ten healthy volunteers (five males and five females) had their biceps brachii, gastrocnemius, and quadriceps muscle examined with strain- and shear wave elastography at three different depths and in regions located above bone and beside bone. Strain ratios were averaged from cine-loops of 10 s length, and shear wave speeds were measured 10 times at each target point. The distance from the skin surface to the centre of each region-of-interest was measured. Measurements were evaluated with descriptive statistics and linear regression. Results: Linear regression showed a significant influence on strain ratio measurements from the reference region-of-interest position, i.e. being above the same structures as the target region-of-interest or not (means: 1.65 and 0.78; (P < 0.001)). For shear wave speeds, there was a significant influence from depth and location above or beside bone (P = 0.011 and P = 0.031). Conclusion: Strain ratio values depend significantly on reference and target region-of-interest being above the same tissue, for instance bone. Strain ratios were not influenced by depth in this study. Shear wave speeds decreased with increasing scanning depth and if there was bone below the region-of-interest.