SummarySeveral studies have investigated the relationship between ultrasound‐measured muscle thickness (MT) and individual muscle cross‐sectional area (CSA) and muscle volume (MV) in extremity and trunk muscles; however, the hamstring muscle has not been studied. The purpose of this study was to examine the relationship between posterior thigh MT by ultrasound and the muscle CSA and MV of the hamstring obtained by magnetic resonance imaging (MRI). Ten young women aged 20–31 had MT measured by ultrasound at three sites on the medial anterior (50% of thigh length; TL) and posterior (50% and 70% of TL) aspects of the thigh. On the same day, a series of continuous muscle CSA along the thigh was measured by MRI. In each slice, the anatomical CSA of the hamstring (biceps femoris, semitendinosus and semimembranosus) and quadriceps muscle was analysed, and the CSAs at 50% and 70% of TL and maximal CSA of the hamstring (CSAmax) were determined. MV was calculated by multiplying CSA by slice thickness. A significant correlation was observed between posterior 50% MT and 50% hamstring CSA (r = 0·848, P = 0·002) and between posterior 70% MT and 70% hamstring CSA (r = 0·679, P = 0·031). Posterior 50% MT (r = 0·732, P = 0·016) and 50% MTxTL (r = 0·873, P = 0·001) were also correlated to hamstring MV. Anterior:posterior 50% thigh MT ratio was correlated to MV ratio of quadriceps and hamstring muscles (r = 0·803, P = 0·005). Our results suggest that posterior thigh MT reflects hamstring muscle CSA and MV. The anterior:posterior MT ratio may serve as a surrogate for MV ratio of quadriceps and hamstring.