Immunogold-silver cytochemistry using a capillary action staining system.

An improved method for immunogold-silver staining is described. Rat monoclonal antibodies to T-lymphocyte surface membrane antigens were employed to demonstrate T-cells and their subsets in frozen sections of mouse spleen and in smears of peripheral blood mononuclear cells. A capillary action staining system was used to perform all stages of the procedure, substantially reducing the technical effort involved, saving reagents, and improving reproducibility and control of batch staining. The use of a light-stable silver enhancement reagent simplified the development of the reaction product. Technical modifications were incorporated to obtain good preservation of tissue morphology when employing this reagent on frozen sections and to stabilize the reaction product. The improved immunostaining procedure was rapid, sensitive, and yielded better staining quality, with excellent contrast and minimal nonspecific precipitation of silver.