Abstract
Most investigations with viruses require reliable methods of quantitation. The best method now available for enumerating infectious virus particles is the plaque technic. When suspensions of animal cells are placed in glass containers, the cells settle to the bottom of the suspending nutrient medium and attach to the glass. Upon incubation and periodic addition to, or replacement of, the nutrient medium the cells will multiply and form a monolayer or sheet of cells. When the cell sheets are complete they are washed to remove virus inhibitors, virus is added, and after a short incubation period to allow for infection the cell sheets are overlayed with a nutrient medium containing a solidifying agent. During subsequent incubation virus is released from the infected cells and spreads to adjacent cells; the areas of infection are localized by the solid overlay medium. Eventually the number of viral particles added to the cell sheet can be determined by vital staining and counting the number of foci of infected cells, each area presumably having resulted from infection by a single virus particle.
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