JoVE Encyclopedia of Experiments
Microbiology
0 views • 3:25 min • September 30th, 2026
Take a multi-well plate containing mammalian epithelial cells.
Prepare serial dilutions of a pseudovirus containing coronavirus spike proteins.
Replace the medium with a diluted viral suspension in each well.
Incubate with occasional rocking to allow viral spike proteins to bind to the host cell receptor.
Remove the medium. Add a medium containing methylcellulose to form a semisolid overlay that restricts viral spread, and incubate.
The virus enters the cell and releases its RNA genome.
The RNA directs the synthesis of new viral RNA and proteins, which assemble into viral particles that exit the cell.
The released viruses infect neighboring cells, producing a clear zone called a plaque.
Remove the overlay and wash the cells.
Add a dye and incubate the plate with shaking.
Wash and dry the wells.
The dye stains the cells, while the plaques remain clear.
Count the visible plaques in each well to determine the viral concentration.
To determine the viral titer, plate Vero E6 cells in six well plates at a seeding density of six times 10 to the fifth cells per well and incubate overnight. The next day, set up a tenfold serial dilution series of the viral stock by adding 900 microliters of medium to se
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