JoVE Encyclopedia of Experiments
Microbiology
0 views • 4:33 min • September 30th, 2026
Take a multi-well plate containing epithelial cell monolayers and add a viral suspension to the wells.
Incubate with occasional rocking to evenly distribute the viruses. Viruses attach to and enter the cells.
In one well, add molten agarose, which solidifies into a semi-solid overlay.
In another well, add a microcrystalline cellulose-based viscous liquid overlay and incubate. Its viscosity restricts viral diffusion, similar to the semi-solid overlay.
Inside the cell, the virus releases its genome and produces new viral RNAs and proteins.
New viruses assemble, exit the cells, and cause cell lysis.
Released viruses infect nearby cells, forming plaques.
Aspirate the liquid overlay and add a fixative to preserve the non-lysed cells.
Discard the fixative and remove the agarose overlay.
Add a dye that stains viable cells, leaving plaques unstained. Then, wash off the excess dye.
The liquid overlay produces comparable plaque numbers to the semi-solid overlay, supporting its use as an alternative in plaque assays.
The day after plating, use an inverted microscope to check the confluency and viability of the cells prior to starting the assay. Ensure that the cells exhibit standard cellular morphology
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