Hello ,
Thank-you very much for sharing the Immunoplaque assay protocol .
In Fact the plaque assay did not work due to some constraint from our part :
1. We did not have the 6-well tissue culture dish but used individual tissue culture plate which we placed on a tray to giggle the plate to evenly distribute the media.
2. The cell monolayer (MDCK cell line ) was not 90 % confluent , so we incubated the plate for 2-3 days . Finally the cells grew but not around the rim of the plate. (See picture)
3. Even so we proceeded with inoculation of virus dilutions (Influenza virus) and overlaid the plate with Noble agar , instead of agarose .
4. We did not have MTT available for staining but used 1% Crystal violet .
Final result : No plaque visible .
Quite discouraging but will try again !!
12/13/2018 6:37:35 PM Reply