Comparative analysis of methodological approaches applied for the simultaneous demonstration of several antigens in immunohistochemical studies

In immunomorphological studies, it is often necessary to visualize several antigens simultaneously within the same histological sections. However in practice, the application of double staining is limited due to methodological complexity. The aim of the current study was to reproduce different methods recommended for multiple immunohistochemical staining of one section and to estimate their advantages and drawbacks. Six different methodological approaches for detection of several antigens in one slide were examined. It was found that the method providing the widest resources for analysis was the fluorescent detection, the most convenient was the application of ready-to-use standardized reagents, and the cheapest method was two-color reaction with DAB-type chromogen.