Background: The aim of the study was to evaluate middle cerebellar peduncle (MCP) asymmetry by diffusion tensor imaging (DTI) according to the dominant hand. Materials and methods: Sixty-four volunteers who met the exclusion and inclusion criteria were prospectively evaluated by DTI. Circular voxels of interest were drawn at the right and left MCP levels for all volunteers. The software automatically calculated the median values of fractional anisotropy (FA), mean diffusivity (MD), and number of fibres (NF). The volunteers were categorised as the right-handed (n = 44) and left-handed (n = 20) groups using the Dellatolas test. The mean values of FA, MD and NF were statistically compared between the two sides and the two groups. Results: In both groups, the mean value of FA for the right MCP was significantly higher than the left MCP (p = 0.0016 and p = 0.0032 for the right-and left-handed groups, respectively). In both groups, no significant difference was observed between the right and left MCPs in terms of MD and NF (p = 0.75, p = 0.69 and p = 0.96, p = 0.46, respectively). There was also no statistically significant difference between the groups for the mean values of FA, MD and NF of each MCP (p = 0.74, p = 0.26 and p = 0.26, respectively for the right MCP and p = 0.78, p = 0.37 and p = 0.54, respectively for the left MCP). Conclusions: There is an asymmetry between the mean FA values of the right and left MCP. However, this asymmetry is independent of the dominant hand. (Folia Morphol 2019; 78, 3: 481-486)