Postembedment staining of complex carbohydrates: Influence of fixation and embedding procedures

In recent years technological advancements have led to improvements in ultrastructural cytochemical methods for localizing and characterizing complex carbohydrates. In particular the introduction of lectins with specific affinities for various sugars and sugar sequences as histochemical probes has increased knowledge concerning the cellular and subcellular distribution of glycoconjugates. Development of nonepoxy‐based embedding materials has provided increased sensitivity compared to the earlier less specific methods and the current lectin methods for localizing sugar moieties. Postembedment staining based on the reactivity of functional groups present in sugars, such as hydroxyl groups, vicinal diol groups, carboxyl groups, and sulfate esters, requires specific conditions for tissue fixation and embedding. The same requirements pertain to staining based on lectin binding. The influence of fixation and embedment using older and newly developed embedding mixtures on the ultrastructural demonstration of complex carbohydrates is considered in this discussion. Fixation with osmium tetroxide and embedment in epoxy resins provides the least sensitive combination for the detection of the reactive groups of complex carbohydrates. The best ultrastructural demonstration of glycoconjugates is achieved when nonosmicated tissues are embedded in nonepoxy resins.