Abstract
Monolayers of human foreskin fibroblast cells were infected with a high input multiplicity of coronavirus 229-E. Virus was allowed to adsorb to the cells for 2 hours at 37°C. After this interval, one series of cell monolayers was covered with Eagle's minimal essential medium with 2% bovine serum warmed to 33°C and incubated in an atmosphere of 5% CO2 at 33°C. Another cell monolayer series was covered with the same medium at 37°C and incubated in an atmosphere of 5% CO2 at 37°C. At appropriate intervals, cultures were switched from 33°C to 37°C and incubated for a total of 24 hours post-inoculation. After incubation, all cultures were harvested for titration of infectious virus or electron microscopy.The monolayers for electron microscopy were fixed in 6% glutaraldehyde, post-fixed in 2% OsO4 in Clark's buffer, dehydrated in an ethanol series, and embedded in Epon 812. Ultra-thin sections were prepared and double stained with lead citrate and uranyl acetate.
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