The aim of this article is to elucidate the ideal sites for botulinum toxin injection by examining the intramuscular nerve distributions in the hamstring muscles. The hamstring muscles, biceps femoris, semitendinosus, and semimembranosus (10 specimens each) were stained by the modified Sihler method. The locations of the muscle origins, nerve entry points, and intramuscular arborized areas were recorded as percentages of the total distance from the line crossing the medial and lateral tibial condyles (0%) to the ischial tuberosity (100%). Intramuscular arborization patterns were observed at 15–30% and 50–60% for the biceps femoris, 25–40% and 60–80% for the semitendinosus, and 20–40% for the semimembranosus. This study suggests that botulinum toxin injection for spasticity of the hamstring muscles should be targeted to specific areas. These areas, where the arborization of intramuscular nerve branches is maximal, are recommended as the most effective and safest points for injection. Clin. Anat. 29:746–751, 2016.
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