Backscattered electron imaging by scanning electron microscopy of regenerating peripheral nerve axons immunostained...

To demonstrate three-dimensional architecture of regenerating axons growing through basal lamina tubes in cryoinjured nerve graft, backscattered electron (BSE) imaging in a scanning electron microscope (SEM) was used to visualize immunostained axons. Regenerating axons immunostained with an antibody against the 200 kD neurofilament protein (RT97) were clearly visualized in BSE images as bright components pursuing an irregular, often spiral course within the basal lamina tubes, and commonly branching within the tubes. The morphology of these structures corresponded closely to that of putative regenerating axons in SEM preparations following application of the potassium hydroxide-collagenase digestion method. The present approach, however, is a considerable improvement on the latter, providing three-dimensional information together with the identification of regenerating axons.