Element Constituents of Rodent Bulbourethral Gland Goblet Granules, Vertebrate Mast Cell Granules and Human Platelet Dense Bodies by Quantitative X-Ray Microanalysis on Fresh Preparations

Abstract

Fresh preparations including fresh frozen dried ultrathin sections, prints and smears and fresh air-dried prints and smears were prepared of the rat, hamster and guinea pig bulbourethral glands (BUG), tree frog tongue, red ear turtule blood and goldfish intestine and adult normal and allergic dermatitis and chronic myelogenous leukemia patient blood. They were observed under a 200 kV transmission electron microscope at acceleration voltages of 200 kV and 80 kV. They were then observed under the analytical electron microscope (AEM, X-650) in a scanning transmission (STEM) mode at an acceleration voltage of 40 kV at specimen currents of 0.2, 1 and 2 nA. Goblet cell granules, mast cell and basophil granules and platelet dense bodies were analyzed under AEM in a STEM mode at an acceleration voltage of 40 kV, specimen current of 1 or 2 nA, counting time of 100 sec. The quantification was made using a program prepared by the standard solution of 20% polyvinylpyrollidone containing different kinds of compounds of 16 different elements. The concentration was expressed in mM/KG dry weight. In the fresh frozen dried ultrathin sections prepared by a new freezing method with a microwave, the areas extending up to 100 ftm deep from the specimen surface were free from ice-crystal artifacts and the limiting membrane of the goblet granules were distinctly recognized surrounding the individual granules. A quantitative X-ray microanalysis (QXMA) revealed that goblet cell granules of the rat, hamster and guinea pig BUG contained high concentrations of sulfur and magnesium and small amounts of calcium and sulfur and magnesium showed close positive correlations in their concentration. Among forty nine mucous granules of the rat BUG cryohxed with microwave, most contained high concentrations of sulfur and magnesium while a few had high concentrations of sulfur and potassium and small amounts of magnesium or calcium. QXMA of mast cell granules of the tree frog tongue, spleen and peritoneum on fresh frozen dried ultrathin sections, fresh frozen dried and fresh air-dried prints, red ear turtle blood basophils on fresh air-dried smears and goldfish small intestine on fresh frozen dried ultrathin sections disclosed high concentrations of sulfur and magnesium in tree frog mast cell and basophil granules, large amounts of zinc in goldfish intestine mast cell granules while basophil granules of the turtle blood contained high concentrations of sulfur and a very small amount of magnesium or calcium. QXMA of human blood platelet dense bodies revealed high concentration of phosphorus and calcium and very small amounts of magnesium in all the persons examined except one AD and one GML patient whose platelet dense bodies contained high concentrations of magnesium. Divalent cations in mast cells are probably in close relation with proteoglycans, those in goblet cell granules with glycoproteins and those in platelet dense bodies bound to nucleotides such as ATP and ADP within the granules respectively. © 1994, JAPAN SOCIETY OF HISTOCHEMISTRY AND CYTOCHEMISTRY. All rights reserved.