OBJECTIVE: Because of their complex topography, long courses, and small diameters, peripheral nerves are challenging structures for radiological diagnostics. However, imaging techniques in the area of peripheral nerve diseases have undergone unexpected development in recent decades. They include MRI and high-resolution sonography (HRS). Yet none of those imaging techniques reaches a resolution comparable to that of histological sections. Fascicles are the smallest discernable structure. Optical coherence tomography (OCT) is the first imaging technique that is able to depict a nerve's ultrastructure at micrometer resolution. In the current study, the authors present an in vivo assessment of human peripheral nerves using OCT. METHODS: OCT measurement was performed in 34 patients with different peripheral nerve pathologies, i.e., nerve compression syndromes. The nerves were examined during surgery after their exposure. Only the sural nerve was twice examined ex vivo. The Thorlabs OCT systems Callisto and Ganymede were used. For intraoperative use, a hand probe was covered with a sterile foil. Different postprocessing imaging techniques were applied and evaluated. In order to highlight certain structures, five texture parameters based on gray-level co-occurrence matrices were calculated according to Haralick. RESULTS: The intraoperative use of OCT is easy and intuitive. Image artifacts are mainly caused by motion and the sterile foil. If the artifacts are kept at a low level, the hyporeflecting bundles of nerve fascicles and their inner parts can be displayed. In the Haralick evaluation, the second angular moment is most suitable to depict the connective tissue. CONCLUSIONS: OCT is a new imaging technique that has shown promise in peripheral nerve surgery for particular questions. Its resolution exceeds that provided by recent radiological possibilities such as MRI and HRS. Since its field of view is relatively small, faster acquisition times would be highly desirable and have already been demonstrated by other groups. Currently, the method resembles an optical biopsy and can be a supplement to intraoperative sonography, giving high-resolution insight into a suspect area that has been located by sonography in advance.