Histochemical observations on the role of ferric chloride in the high-iron diamine technique for localizing sulphated mucosubstances.

Abstract

The binding of ferric ions to tissue sites, other than those containing sulphated mucosubstances, in sections subjected to the high iron diamine technique was followed by the Prussian blue reaction in order to throw new light on the role of ferric chloride in the high-iron diamine dye bath. From experiments involving enzyme digestions in particular, evidence was obtained that ferric ions bind to ribonucleic acid (in chief cells of the rabbit stomach), deoxyribonucleic acid (in nuclear chromatin granules), and, under certain conditions, to sialic acid residues (e.g. in mucous acini of the mouse sublingual gland) and protein carboxyl groups (in smooth muscle cells) as well. With a few exceptions, the binding of ferric ions to nucleic acids was not affected by changes in the ferric chloride concentration, pH or magnesium chloride concentration in the dye bath; the bond thus formed was very stable. It is possible that the initial linkage is not an electrostatic one. Under all the conditions investigated, it was found that the diamine complexes have a greater affinity for sulphated mucosubstances than ferric ions, but ferric chloride, by lowering the pH of the dye bath, excludes the carboxyl groups from reacting with the positively-charged diamine polymer molecules. It is possible that a high concentration of ferric ions in the high-iron diamine dye bath inhibits the binding of some diamine complexes to nuclei and gastric chief cells, i.e. sites where no sulphated mucosaccharides are present, although this conclusion needs further substantiation.