The simultaneous observations of immunofluorescence and cellular morphology in viral infections: a new procedure

A simple method for comparative observations of immunofluorescence and cellular morphology has been developed in the investigation of Epstein-Barr virus (EBV) infection and oncogenesis. When cells were stained for immunofluorescence of EBV-determined nuclear antigen, early antigens, or the viral capsid antigen, followed by treatment with hematoxylin, viral marker expression and cellular morphology could be simultaneously observed in individual cells by simply changing ultraviolet illumination to visible light. In our double-staining method, hematoxylin was not counteractive to specific immunofluorescence, and pretreatment of cells with antibodies did not affect hematoxylin staining. This method may possibly be more widely applied in virology and other fields.