Electron microscopy of viruses and virus-cell interactions.

Electron microscopy is a powerful tool to visualize viruses in diagnostic as well as in research settings for investigating viral structure and virus-cell interactions. Here, a simple but efficient method is described for demonstrating viruses by negative staining, and its limit is discussed. A prerequisite to obtain reliable information on virus-cell interactions is excellent preservation of cellular and viral ultrastructure. The crux is that during fixation and embedding, by applying conventional protocols about 50% of the lipids are lost, which results in loss of integrity of cell membranes. To achieve good preservation of cellular architectures, good contrast, and both high spatial and temporal resolution, methods for freezing, freeze-substitution, and freeze-etching are described and their applicability discussed mostly taking complicated built herpes viruses as examples.