Ultrastructural detection of calcium and magnesium in the chromatoid body of mouse spermatids by electron spectroscopic imaging and electron energy loss spectroscopy.

We studied at the ultrastructural level the presence of calcium and magnesium in the chromatoid body (CB) in mouse spermatids. In addition, the presence of these two cations was also examined in the nucleolus. By electron spectroscopic imaging (ESI) and electron energy loss spectroscopic (EELS) analyses on glutaraldehyde/pyroantimonate-fixed material, we showed the association of pyroantimonate precipitate granules containing calcium and/or magnesium with the CB. The granules in both active (primary spermatocytes) and inactive (spermatids) nucleoli contain mainly calcium. Our results confirm that although the pyroantimonate technique used alone suffers from a lack of specificity, it can be useful for in situ immobilization of different cations that are further resolved with methods of elemental analysis. Without pyroantimonate fixation, only very few spots containing calcium can be localized by ESI and EELS.