Tendon properties impact human locomotion, influencing sports performance, and injury prevention. Hamstrings play a crucial role in sprinting, particularly the biceps femoris long head (BFlh), which is prone to frequent injuries. It remains uncertain if BFlh exhibits distinct mechanical properties compared to other hamstring muscles. This study utilized free‐hand three‐dimensional ultrasound to assess morphological and mechanical properties of distal hamstrings tendons in 15 men. Scans were taken in prone position, with hip and knee extended, at rest and during 20%, 40%, 60%, and 80% of maximal voluntary isometric contraction of the knee flexors. Tendon length, volume, cross‐sectional area (CSA), and anteroposterior (AP) and mediolateral (ML) widths were quantified at three locations. Longitudinal and transverse deformations, stiffness, strain, and stress were estimated. The ST had the greatest tendon strain and the lowest stiffness as well as the highest CSA and AP and ML width strain compared to other tendons. Biceps femoris short head (BFsh) exhibited the least strain, AP and ML deformation. Further, BFlh displayed the highest stiffness and stress, and BFsh had the lowest stress. Additionally, deformation varied by region, with the proximal site showing generally the lowest CSA strain. Distal tendon mechanical properties differed among the hamstring muscles during isometric knee flexions. In contrast to other bi‐articular hamstrings, the BFlh high stiffness and stress may result in greater energy absorption by its muscle fascicles, rather than the distal tendon, during late swing in sprinting. This could partly account for the increased incidence of hamstring injuries in this muscle.