Visualization of viral assembly in the infected cell.
The study of the virus life cycle in infected cells is a methodological challenge due to the small size and diversity of the viral components. Recent developments on preservation of fine structure and molecular localization have provided a group of powerful methods with wide applications in cell biology and virology. Among the different electron microscopy (EM) techniques available to visualize viral assembly at the intracellular level, we will focus on conventional ultrathin sections, cryosections, and freeze-substitution. For obtaining molecular information associated to ultrastructure we have now a group of methods to detect viral proteins (immunogold labeling), as well as the viral genome, through the different techniques for detection of nucleic acids (the enzyme-gold approach, in situ hybridization, and elemental mapping). We will illustrate the applications of these methods with examples of viruses that exhibit different levels of structural complexity. These new approaches help to detect and identify viruses in clinical samples and to characterize the virus life cycle and the cellular components involved, to obtain data that could help for a therapeutic intervention, and to characterize virus-like particles that can be the basis of new and safe vaccines.
Risco
C
C
Department of Macromolecular Structure, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, Spain. crisco@cnb.uam.es
Carrascosa
J L
JL
D000818:Animals; D020285:Cryoelectron Microscopy; D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D012367:RNA, Viral / Q000032:analysis; D019065:Virus Assembly; D014780:Viruses / Q000648:ultrastructure
D004279:DNA, Viral; D012367:RNA, Viral