Title: Enzymatic Digestion is also an Effective Antigenicity-Restoring Method for Immunohistochemistry at the Electron Microscope (EM) Level
Abstract: Immunohistological study using EM technique offers several advantages over light microscopy (LM) such as unequivocal identification of cell type by morphology and accurate subcellular localization of antigenic molecules of interest. These are mainly attributed to fixation with strong cross-linking reagents and embedding in resin-based media. However, they also are associated with a major drawback -- greatly reduced antigenicity or completely abolished accessibility to tissue antigen due to extensive cross-linking and the hydrophobicity of the resin. As a result, the success rate of immunolabeling of routinely processed tissue for EM is far lower than that for LM using paraffin sections. Yet the process is far more laborious. To circumvent this problem, various etching or de-plasticizing methods have been introduced. However, most of them invariably lead to deterioration of the tissue morphology. A trade-off between morphology and antigenicity is generally considered necessary. Here we demonstrate that a brief enzymatic digestion prior to immunolabeling the routinely processed tissue samples for EM greatly improves the success rate of the immunostaining without noticeable effect on morphology. Our results suggest that enzymatic digestion is an effective antigenicity-restoring method for EM.
Publication Year: 1996
Publication Date: 1996-08-11
Language: en
Type: article
Indexed In: ['crossref']
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