B-mode ultrasound (US) is often used to noninvasively measure skeletal muscle architecture, which contains human intent information. Extracted features from B-mode images can help improve closed-loop human–robotic interaction control when using rehabilitation/assistive devices. The traditional manual approach to inferring the muscle structural features from US images is laborious, time-consuming, and subjective among different investigators. This paper proposes a clustering-based detection method that can mimic a well-trained human expert in identifying fascicle and aponeurosis and, therefore, compute the pennation angle. The clustering-based architecture assumes that muscle fibers have tubular characteristics. It is robust for low-frequency image streams. We compared the proposed algorithm to two mature benchmark techniques: UltraTrack and ImageJ. The performance of the proposed approach showed higher accuracy in our dataset (frame frequency is 20 Hz), that is, similar to the human expert. The proposed method shows promising potential in automatic muscle fascicle orientation detection to facilitate implementations in biomechanics modeling, rehabilitation robot control design, and neuromuscular disease diagnosis with low-frequency data stream.