The influence of embedding media and fixation on the post-embedment ultrastructural demonstration of complex carbohydrates. I. Morphology and periodic acid-thiocarbohydrazide-silver proteinate staining of vicinal diols.

The influence of fixation and embedding medium on the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) staining reactivity in the mouse intestine was studied. It was found that the combination of osmium tetroxide and epoxy resins was the least sensitive for the demonstration of complex carbohydrate with the PA-TCH-SP method. Post-osmication reduced, but did not abolish, PA-TCH-SP reactivity (except for the Golgi complex) when non-epoxy resins were used. The staining pattern of a particular organelle differed depending on the embedding medium used. Golgi cisternae exhibited the most intense PA-TCH-SP reactivity in non-osmicated tissues embedded in non-epoxy resins. Post-osmication of tissues was required to reveal the fine structure of the glycocalyx as well as to preserve the fine structure of tissues embedded in styrene-methacrylate and styrene-Rigolac 2004. The choice of fixation procedures and embedding media in a given study should be governed primarily by the sites of interest.