Functional MRI (fMRI) enables evaluation of language cortical organization and plays a central role in surgical planning. Diffusion Tensor Imaging (DTI) or Tractography, allows evaluation of the white matter fibers involved in language. Unlike fMRI, DTI does not rely on the patient's cooperation. In monolinguals, there is a significant correlation between the lateralization of language on fMRI and on DTI. Our objective is to delineate the arcuate fasciculus (AF) in right- and left-handed trilinguals and determine if the AF laterality on DTI is correlated to language lateralization on fMRI. 15 right and 15 left-handed trilingual volunteers underwent fMRI and DTI. Laterality Index was determined on fMRI (fMRI-LI). Mean Diffusivity, Fractional Anisotropy (FA), Number of Fibers, Fiber Length, Fiber Volume and Laterality Index (DTI-LI) of the AF were calculated on DTI. 28 of the 30 subjects presented a bilateral AF. Most subjects (52%) were found to have a bilateral language lateralization of the AF on DTI. Only 4 subjects had bilateral lateralization of language on fMRI. The right AF demonstrated lower diffusivity than the left AF in the total participants, the right-handed, and the left-handed subjects. FA, Volume and Length of the AF were not significantly different between the two hemispheres. No correlation was found between the DTI-LI of the AF and the fMRI-LI. A prominent role of the right AF and a bilateral structural organization of the AF was present in our multilingual population regardless of their handedness. While in prior studies DTI was able to determine language lateralization in monolingual subjects, this was not possible in trilingual highly educated subjects.