Sensory and motor fiber differentiation with Karnovsky staining

We examined four acethylthiocholine methods based on Karnovsky's procedure—two fast-acting requiring 1 hour and two slow-acting requiring 24 hours. We compared these with our modification, which requires less than an hour and is simple to use. Rabbit sciatic nerves and spinal cords were used to compare methods. Our modification showed clearer differentiation than other fast-acting methods and staining identical to slow-acting methods. In blind examination of radial nerve specimens stained with our method, motor and sensory fascicles were correctly identified, showing sensitivity and specificity of 100%. In 12 clinical cases, our method produced staining in the proximal stump as long as 16 months after injury and in the distal stump as long as 5 days after injury. In 10 of 12 patients, this staining helped in aligning motor fascicles to motor fascicles and sensory fascicles to sensory fascicles.