Improved post-embedding immunocytochemistry of myelinated nervous tissue for electron microscopy

Abstract

The particularly high lipid content of normal mature adult myelin sheaths, together with the light fixation protocols usually necessary to retain antigenicity, combine to make white matter nervous tissue an especially problematical subject for post-embedding immuno-electron microscopy using modern acrylic resins. Fixation and infiltration modifications to standard processing schedules for Lowicryl were found to greatly improve the embedding and therefore the resulting morphology. This in turn improved the signal to noise ratio by reducing the high non-specific backgrounds usually found in poorly infiltrated areas. Using Lowicryl HM20, we have been able to obtain satisfactory immunostaining for myelin basic protein with good retention of structural integrity in the myelin of both normal and lesioned adult cortico–spinal tract.