The use of immunogold-silver staining (IGSS) to enhance label penetration and Localization for immunocytochemistry or in situ hybridization utilizing a variety of metallic salts has been documented. In this morphological study, the effects of silver acetate, silver lactate and silver nitrate were evaluated for immunogold-labeling of a trial natriuretic peptides (ANP) in rat right atria.Eight Wistar Kyoto retired breeders were sedated with pentobarbital, perfused with either 4% paraformaldehyde (LM) or Karnovsky's fixative (EM), and right atria were dissected, processed, embedded in paraffin or epon, respectively and sectioned according to conventional methods. For light microscopy, an indirect IGSS method according to Hacker (3) was performed. Paraffin sections on glass slides were washed in ddH2O, immersed in Lugol's iodine, washed in ddH2O and treated with 2.5% aqueous sodium thiosulfate for 20 sec. After additional washes in ddH2O and TBS-0.1% fish gelatin, 10% normal goat serum (PBS with 1% BSA) was applied for 20 min before an overnight incubation at 4°C with a polyclonal α-ANP primary antibody (Peninsula Labs, 1:1000 in TBS/BSA).
Electrophoretic identification of muscle proteins in several puffer species with Coomassie blue/silver staining