Semiquantitative evaluation of histochemically detectable calcium binding in mammalian brain slices

A method is described for the electron histochemical demonstration of Ca in the central nervous system, based on fixation on Ca-containing paraformaldehyde solution and the subsequent complexing of Ca by ammonium oxalate. The method resulted in highly electron-dense deposits, with good ultrastructural preservation. The Ca content of the deposits was proved by physico-chemical analysis. The high electron density permitted the counting of deposits and thereby an estimation of their numerical density, via planimetry of electron micrographs. Since pre- and postsynaptic localizations could be distinguished on the basis of ultrastructure, this procedure is regarded as a unique semiquantitative method for estimation of the tissue Ca binding of mammalian brain slices.