Muscle morphology is an important contributor to hamstring muscle injury and malfunction. The aim of this study was to examine if hamstring muscle-tendon lengths differ between various measurement methods as well as if passive length changes differ between individual hamstrings. The lengths of biceps femoris long head (BFlh), semimembranosus (SM), and semitendinosus (ST) of 12 healthy males were determined using three methods: Firstly, by identifying the muscle attachments using ultrasound (US) and then measuring the distance on the skin using a flexible ultrasound tape (TAPE-US). Secondly, by scanning each muscle using extended-field-of view US (EFOV-US) and, thirdly, by estimating length using modelling equations (MODEL). Measurements were performed with the participant relaxed at six combinations of hip (0°, 90°) and knee (0°, 45°, and 90°) flexion angles. The MODEL method showed greater BFlh and SM lengths as well as changes in length than US methods. EFOV-US showed greater ST and SM lengths than TAPE-US (p < 0.05). SM length change across all joint positions was greater than BFlh and ST (p < 0.05). Hamstring length predicted using regression equations is greater compared with those measured using US-based methods. The EFOV-US method yielded greater ST and SM length than the TAPE-US method. SM showed the highest change in length at different hip and knee joint positions.