Histochemical studies on the stainability of alum hematoxylin and its staining mechanism
Kaneko, K.; Akita, M.; Murata, E.; Fujita, K.
Kaibogaku Zasshi. Journal of Anatomy 60(4): 290-301
1985
ISSN/ISBN: 0022-7722 PMID: 2420144 Document Number: 250389
The multiformity in the stainability of alum hematoxylin was studied at various pH values using rat and mouse tissues and model substances in combination with enzyme digestion and chemical modification procedures. Alum hematoxylin did not show stainability at strongly acidic solutions of approx. pH 1, however cell nuclei began to be stained at about pH 2. As acidic solutions below pH 4, alum hematoxylin partially stained nuclei acids in the form of basic complex. However, since is also stained basic amino acids such as arginine and histidine, the concept is presented that it stains both basic nucleoproteins and nuclei acids. At pH 6-8, alum hematoxylin stained basic amino acids in the form of an acidic complex, thus it appeared to stain only basic nucleoproteins. At pH 3-4, alum hematoxylin stained the cartilage matrix; it also stained acidic mucins in goblet cells of the small and large intestine. In these cases, the basic complex of alum hematoxylin bound with sulfonic groups of chondroitin sulfates in the cartilage matrix and with carboxylic groups of sialic acids in goblet cells of the small and large intestines, forming ionic linkages. At pH 6, alum hematoxylin stained glycogen granules in the liver cells and mucins in mucous neck cells of the stomach, forming a hydrogen bond. At pH 6-8, alum hematoxylin stained elastic fibers. At pH 5 and above, and in alkaline solution, alum hematoxylin did not shown stainability as a basic complex but worked as an acidic complex.