DUAL STAINING OF LYMPHOCYTE MEMBRANE-ANTIGENS WITH COLLOIDAL GOLD ANDBIOTINYLATED HORSERADISH-PEROXIDASE

Double immunoenzymatic labelling procedures for the localization of an tigens on cells in tissue sections using horseradish peroxidase (HRP) and alkaline phosphatase have been described previously, but mainly fo r detecting antigens on different cells. With this type of staining wh en two antigens are present on the same cell, an optimal colour combin ation that shows a high contrast between the basic colour of each enzy me substrate product is difficult to achieve and the interpretation of their mixed colour intermediate is subjective. We present a method fo r the simultaneous demonstration of two antigens on the same cell. The method can be used to label either single cells in suspension, or cel ls in paraffin fixed tissue, using a combination of a particulate labe l, colloidal immunogold-silver, and an enzymatic label HRP-DAB. The me thod is easy to perform and utilises commerically available staining k its.