The purpose of this study is to prospectively investigate the reference values of masseter and temporal muscle thicknesses by ultrasonography and muscle hardness values by shear wave elastography in healthy adults. The sample of the study consisted of a total of 160 healthy individuals aged between 18 and 59, including 80 women and 80 men. By examining the right and left sides of each participant, thickness and hardness values were obtained for 320 masseter muscles and 320 temporal muscles in total. The mean masseter muscle thickness was found to be 1.09 cm at rest and 1.40 cm in contraction. The mean temporal muscle thickness was found to be 0.88 cm at rest and 0.98 cm in contraction. The thickness values of the masseter and temporal muscles were significantly greater in the male participants than in the female participants (P < .001). While there were significant differences between the right and left masseter muscle thickness values at rest and in contraction, the values of the temporal muscles did not show a significant difference between the sides. While the resting hardness (rSWE) of the masseter muscle was transversally 6.91 kPa and longitudinally 8.49 kPa, these values in contraction (cSWE) were found, respectively, 31.40 and 35.65 kPa. The median temporal muscle hardness values were 8.84 kPa at rest and 20.43 kPa in contraction. Masseter and temporal muscle hardness values at rest and in contraction were significantly higher among the male participants compared to the female participants (P < .001). In this study, reference values for the thickness and hardness of the masseter and temporal muscles are reported. Knowing these values will make it easier to assess pain in the masseter and temporal muscles and determine the diagnosis and prognosis of masticatory muscle pathologies by allowing the morphological and functional assessments of these muscles, and it will identify ranges for reference parameters.