Electron microscopy of viruses

Abstract

Electron microscopy is widely used in virology because viruses are generally too small for a direct inspection by light microscopy. Analysis of virus morphology is necessary in many circumstances, e.g., for the diagnosis of a virus in particular clinical situations or the analysis of virus entry and assembly. Moreover, quality control of virus particle integrity is required if a virus is propagated in cell culture, particularly if the virus genome has changed. In most cases already the basic methodology for transmission electron microscopy, i.e., negative staining and ultrathin sectioning, is sufficient to give relevant information on virus ultrastructure. This chapter gives detailed information on the principles of these basic methodologies and provides simple but reliable protocols for a quick start. Moreover, the description of standard protocols for negative staining and ultrathin sectioning are supplemented by protocols on immuno-negative staining and rapid ultrathin sectioning. Finally, principles of methods for an extended ultrastructural research using more elaborate techniques, such as cryotechniques or methods to reveal the three-dimensional virus architecture, are briefly reviewed.