The palmaris brevis (PB) is the only muscle frequently innervated by the superficial branch of the ulnar nerve. It is clinically involved in patients with sensory impairment of the ulnar nerve sparing the deep branch as seen in mild cases of cyclist or crutch palsy. It is also involved in palmaris brevis spasm syndrome. A technique is described that makes it possible to localize the PB distinctly from its neighbors by electromyographic (EMG) pin examination. Methods include prospective (1) anatomic measurement in 10 cadavers and (2) EMG pin examination in 12 normal subjects measuring rise time (ms) and RMS voltage output (microV). Results showed (1) the area of the PB in elderly cadavers was 2.1+/-0.7 x 2.1+/-0.3 cm with a thickness of 0.2+/-.01 cm and (2) in the live subjects, the muscle could be located with identification of motor unit potentials having a rise time <1 ms 100% of the time. In conclusion, the PB can be localized for EMG analysis by using an approach parallel to surface of the palm. Clinically, it could be a useful complement to diagnose neuropathy involving the superficial fibers of the ulnar nerve.