Immunofluorescence microscopy and immunogold EM for investigating fungal infection of plants.

Fungi are a diverse group of eukaryotic organisms whose activities are intricately linked to the lives of human beings. Their involvement in plant productivity, as agents of human diseases, as sources of medicines and enzymes and as model experimental organisms has necessitated the development of sensitive and specific techniques for tracking the organisms and their protein products. Techniques employing highly specific monoclonal antibodies have allowed the visualization of fungi in their natural environments and have facilitated the study of their antigens at the subcellular level. Here, we describe three such techniques, immunofluorescence (IF), immuno-enzymatic staining (IES) and immunoelectron microscopy (IEM), that have found widespread applicability in studies of fungal biology, and which can also be adapted for use in the study of other eukaryotic organisms. Results from the IF and IES procedures can be obtained within 4-5 h. Sample preparation for IEM takes approximately 4 days. Gold labeling and visualization of samples can be completed within 4 h.