Direct observation of the budding and fusion of an enveloped virus by video microscopy of viable cells

Video-enhanced microscopy and digital image processing were used to observe the assembly, budding, and fusion of Respiratory Syncytial virus. Viral filaments were seen to bud from the plasma membrane of viable infected cells to a final length of 5-10 micron with an average speed of elongation of 110-250 nm/s. The rapidity of viral assembly and its synchronous occurrence (leading to the production of several viral particles per minute from the same surface domain) suggests a directed process of recruitment of viral components to an area selected for virus maturation. Virions were also seen to adsorb to the cell surface, and to fuse with the plasma membrane. These are the first real time observations of viral morphogenesis and penetration which are crucial events in the infectious cycle of enveloped viruses.