Application of Heidenhain's iron hematoxylin staining to epon-embedded tissues.

Abstract

A method for the application of Heidenhain's iron hematoxylin staining to epon-embedded tissue sections is described.Various tissues of the mouse were fixed with glutaraldehyde followed by osmium tetroxide, embedded in epon-resin, and cut into semi-thin sections about 1μ thick. The sections were affixed to the slide surface by heating on a hot-plate.Heidenhain's iron hematoxylin solution was prepared according to ROMEIS' recipe. The staining procedure which has been utilized for paraffin and celloidin sections was applied to the epon-sections with slight modification, e. g., heating the utilized solutions. The embedding medium was not removed for the staining.The results of the present staining show that the connective tissue fibers, nuclear chromatins and nucleoli were commonly stained in dark shades of black, while the cytoplasm and nuclear sap showed light shades of yellowish brown. The keratinized layer of stratified squamous epithelium, glycogen areas of hepatic cell, goblet cell mucus, Paneth cell granules and the striated border of intestinal epithelium were selectively emphasized in dark shades. On the other hand, the sarcoplasm of muscle fibers, myelin sheaths and erythrocytes appeared in light shades. Under mounting with Canada balsam the stain remains stable for at least a year.The present method is useful especially in tissues containing a cosiderable amount of connective tissue fibers but is valid also as a general stain for all epon-embedded tissues.