Focus measure (FM) plots for 7 different metrics in three different anatomical regions of the mouse during in vivo imaging of (a) brain, (b) liver, and (c) kidney/spleen. The global minima of the focus measure score represent the calibrated speed of sound. Reconstructed images at different speed of sound values for the respective regions are shown in (d–f), where the first and second columns correspond, respectively, to the speed of sound in water (at 34 °C) and the speed of sound manually fitted.