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 tra nscardially 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 gl ial cells markers glial fibrillary acidic protein and vimentin, and se rum albumin having a capacity for binding of zinc and copper were dete cted by immunofluoresence histochemistry. The sections were examined f or the presence of both metals and proteins in a microscope equipped f or both light and epifluorescence microscopy. The reaction product of the autometallographic reaction did not interfere with immunofluoresce nce histochemistry, and vise versa. The method can be used for co-loca lization of zinc and copper with specific cell-marker proteins. Furthe rmore, the method can be used for simultaneous intracellular visualiza tion of these metals and their respective metal-binding proteins.