Scanning electron microscopic observation of immunohistochemically specified stromal cells following the NaOH maceration method

The present study introduces a method combining immunohistochemistry and a chemical digestion technique applied to scanning electron microscopy (SEM) observation of chemically characterized stromal cells in the myenteric layer of the monkey small intestine. The whole-mount preparation containing the myenteric layer was treated immunohistochemically using small colloidal gold and silver enhancements for detecting a certain type of stromal cell, followed by an alkali maceration method to remove extracellular matrices covering the immunohistochemically stained cells. This method permitted direct SEM observation of the stained stromal cells. Secondary and backscatter electron images clearly visualized the ultrastructure of the immunohistochemically-stained cells and the spatial distribution of the reaction products, respectively. It is suggested that the present method is useful for demonstrating the three-dimensional organization of the chemically specified stromal cells.