Methods in Molecular Biology Volume 793, 2011, pp 23-59
Date: 14 Aug 2011

Degenerating Processes Identified by Electron Microscopic Immunocytochemical Methods

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Abstract

The application of electron microscopic immunolabeling techniques to the identification and analysis of degenerating processes in neural tissue has greatly enhanced the ability of researchers to examine apoptosis and other degenerative disease mechanisms. This is particularly true for the early stages of such mechanisms. Traditionally, degenerating processes could only be identified at the ultrastructural level after significant cellular atrophy had occurred, when subcellular detail was obscured and synaptic relationships altered. Using immunocytochemical labeling procedures, degenerating neural and glial processes are first identified through the use of antibodies directed against a variety of degenerative markers, such as proapoptotic effectors (i.e., cytoplasmic cytochrome c), pathological components (i.e., beta amyloid deposits), or inflammatory agents (i.e., Iba1). Both the subcellular distribution of the marker within the process and the relationship of the labeled process to surrounding elements can then be carefully characterized. The information obtained can be further refined through the use of dual immunolabeling, which can provide additional data on the phenotype of the degenerating process and inputs to the process.