The following method has been tried and true for me to dissect single 
myelinated nerve fibers for microscopic examination.

Portion of an unfixed sural nerve biopsy (approx. 0.5 cm) is needed for nerve 
fiber teasing.

Procedure:
1. Nerve is placed into 1% aqueous osmic acid (osmium tetroxide) for 24 hours.
2. Wash in DH2O.
3. Place nerve into 15% aqueous glycerol for 1 hour.
4. Place nerve into 45% aqueous glycerol for 1 hour and then into 60% aqueous 
glycerol over a period of 2 days.
5. Place in 66% aqueous glycerol for teasing. 
6. Proceed with nerve fiber teasing. Nerve is to be stored in 66% glycerol.

Reference:
Bradley W.G.: Disorders of Peripheral Nerves, 1974.



Eric C. Kellar
University of Pittsburgh Medical Center