The functional role of the biceps femoris short head (BFsh) remains unclear. Clarifying the functional role of each biceps femoris head may provide useful insights into the reduction of biceps femoris long head (BFlh) injuries. This study aimed to clarify whether the passive tension in the BFlh would change with BFsh detachment using cadavers. The shear modulus of the BFlh and BFsh was measured using ultrasonic shear wave elastography as an index of passive tension in three tissue processing conditions (intact, removal of all tissues from skin to deep fascia, and BFsh detachment) in four limb positions, with hip [H0°, H90°] and knee [K0°, K90°] joint angles under each tissue processing condition. The measurement site was the distal 30 % of the line connecting the sciatic tuberosity (100 %) and fibular head (0 %). Three-way analysis of variance was conducted with muscles, tissue processing, and positions as factors, which revealed a significant interaction (P < 0.01). The post-test results indicated that the BFlh was significantly higher than the BFsh at H90° and K0° before tissue processing;​ however, no difference was observed between the muscles after skin and deep fascia removal. After BFsh detachment, the shear modulus of the BFsh decreased, whereas that of the BFlh significantly increased (P < 0.01), suggesting that the BFsh might be involved in the passive tension reduction of the BFlh in a lengthened position, as the shear modulus of the BFlh increased after detachment.