SIMULTANEOUS APPLICATION OF TIMM SULFIDE SILVER METHOD AND IMMUNOFLUORESCENCE HISTOCHEMISTRY

This study presents a method for simultaneous detection of metal ions of zinc and copper and proteins in single brain sections. Rat were transcardially perfused with sodium sulphide and subsequently fixed with formalin. Zinc and copper were demonstrated by Timm's sulphide silver staining method using physical development (autometallography). The glial cells markers glial fibrillary acidic protein and vimentin, and serum albumin having a capacity for binding of zinc and copper were detected by immunofluoresence histochemistry. The sections were examined for the presence of both metals and proteins in a microscope equipped for both light and epifluorescence microscopy. The reaction product of the autometallographic reaction did not interfere with immunofluorescence histochemistry, and vise versa. The method can be used for co-localization of zinc and copper with specific cell-marker proteins. Furthermore, the method can be used for simultaneous intracellular visualization of these metals and their respective metal-binding proteins.