The effect of topical glycerol application on normal and previously injured saphenous nerves was tested in 20 Sprague-Dawley rats. Anhydrous glycerol treatment of five normal nerves showed a rapid loss of C-fiber conduction within 5 minutes of application, while after 10 to 30 minutes, a complete conduction blockade in all fiber types was produced. The effect of anhydrous glycerol on both spontaneous firing from the neuroma and impulse propagation within the nerve was examined in 11 rats that had undergone saphenous neurotomy 7 days previously. In these animals, cessation of spontaneous action potential production from the neuroma was the earliest electrophysiological change noted, followed by loss first of C-fiber, then of A-fiber conduction. Graded concentrations of glycerol (25%, 50%, 75%, and 100%) were used in four rats with saphenous neuromas, which allowed selective blockade of spontaneous action potential generation and C-fiber conduction with minimal effect on A-fibers. The neurophysiological mechanism of glycerol neurolysis appears to be a nonspecific conduction blockade of large and small fibers, which is established within minutes of its application. Spontaneous firing within damaged axons, which may play a role in a variety of cranial and peripheral nerve sensorimotor syndromes, is also exquisitely sensitive to glycerol application.