SIMULTANEOUS DEMONSTRATION OF 2 ANTIGENS WITH IMMUNOGOLD-SILVER STAINING AND IMMUNOENZYMATIC LABELING

A novel technique for independent and simultaneous labeling of two ant igens expressed on individual cells (referred to as mixed labeling) is presented. The staining procedure combined three-step (streptavidin-b iotin) immunogold-silver staining with three-step immunoenzymatic labe ling. To ensure both high specificity and high sensitivity, particular emphasis was placed on designing a protocol that avoids immunological crossreactivity between the antibody reagents and overlapping of the final color products. Two examples for usage of this mixed labeling te chnique are described: lymphocyte subpopulations were identified in in flammatory lesions of human skin and infected host cells were characte rized in the skin of mice infected with the obligatory intracellular p arasite Leishmania major, a cause of human cutaneous leishmaniasis.