Teased-fiber technique for peripheral myelinated nerves: methodology and interpretation

Abstract

Teased-fiber technique is the best approach for studying peripheral myelinated nerve fibers in their continuity. It enables the assessment of size of myelin segments formed by Schwann cells and characterization of pathologic changes affecting the internodia, the paranodal regions, and the invested axons. Fiber teasing is performed on prestained proximodistally oriented portions of peripheral nerves. Specimens about 10 mm long are stained for 24-48 hours in Sudan black and then transferred to glycerin, where, using a pair of fine forceps and a stereomicroscope, they are separated into smaller fiber bundles from which single fibers are isolated. The work is performed on a glass slide with an adhesive surface (albuminized or "superfrost"), on which the fibers are placed in strict proximodistal orientation. Following drying in an oven, the slides are mounted with glycerin-gelatine (same as used for frozen sections). The changes, when present, can usually be recognized during the preparation, but fibers are reexamined and changes confirmed in mounted slides. Photographic reconstruction of the fibers facilitates their assessment and enables the documentation of findings. The teased-fiber technique is auxiliary to histopathology, and to limit the workload and save costs, it can be performed on only a few specimens selected for better characterization of changes recognized or suspected in tissue sections. In particular, segmental demyelination and early stages of Wallerian or secondary axonal degeneration can be recognized in teased fibers. Segmental demyelination is characterized by loss of fully myelinated segments and their replacement by newly formed short and thin segments, remyelinating the preserved axon. The early stage of secondary axonal degeneration is recognized by formation of ovoidal fiber fragments in the midinternodal region.