Impedance-Based Cellular Assay to Measure Multiplicity of Infection of Influenza Virus

To determine the correlation between the CIT₅₀ values and the multiplicity of infection, after culturing 3 x 10⁴ freshly split MDCK cells into each wall of the electronic microtiter plate for 24 hours, wash the cells two times with 100 microliters of fresh virus propagation medium per well per wash, and use a single channel pipette to add 100 microliters of viral suspension to each well.

When all of the virus dilutions have been added, gently load the plate into the cradle pocket of the instrument at 35 degrees Celsius, and begin monitoring the cell impedance every 15 minutes for at least 100 hours, as demonstrated.

After two cycles of measurements, click to "Pause" the apparatus and remove the E-plate from the cradle. Add 100 microliters of virus propagation medium supplemented with TPCK-trypsin to each well, and return the E-plate plate into the cradle pocket. Then, start the analysis.