Ultrasound imaging of peripheral nerves is increasingly used in the clinic for a wide range of applications. Although yet unapplied for experimental neuroscience, it also has potential value in this research area. This study explores the feasibility, possibilities and limitations of this technique in rabbits, with special focus on peripheral nerve regeneration after trauma. The peroneal nerve of 25 New Zealand White rabbits was imaged at varying time intervals after a crush lesion. The ultrasonic appearance of the nerve was determined, and recordings were validated with in vivo anatomy. Nerve swelling at the lesion site was estimated from ultrasound images and compared with anatomical parameters. The peroneal nerve could reliably be identified in all animals, and its course and anatomical variations agreed perfectly with anatomy. Nerve diameters from ultrasound were related to in vivo diameters (p < 0.001, R(2) = 77%), although the prediction interval was rather wide. Nerve thickenings could be visualized and preliminary results indicate that ultrasound can differentiate between neuroma formation and external nerve thickening. The value of the technique for experimental neuroscience is discussed. We conclude that ultrasound imaging of the rabbit peroneal nerve is feasible and that it is a promising tool for different research areas within the field of experimental neuroscience.