Cellular Impedance-Based Survival Kinetics Assay to Assess Virus Survival
Transcription
Viruses bind to the host-cell receptor and internalize into an endosome. The endosomal membrane fuses with the viral membrane, releasing viral contents inside the cell.
Upon reaching the nucleus, the viral genome transcribes into mRNA and later translates to form viral proteins. The viral components enter the cytoplasm and assemble into virions, which pinch out of the cell and cause cytopathic effects ― changes in the host cell's morphology, and death.
To assess virus survival kinetics by measuring host-cell impedance, take a multiwell plate coated with impedance-measuring microelectrodes at its bottom. Add a host-cell suspension to the wells, and incubate to allow the cells to adhere to the electrode surface, and grow.
Next, take saline pre-treated viral suspensions with different viral loads. Saline treatment supplies the virus with chloride, which produces hypochlorous acid that kills it ― reducing the viral infectivity. Add these viral suspensions into their respective wells.
Measure the cellular impedance ― a phenomenon of opposing the flow of electric current through the electrodes ― by the adherent cells.
Viruses infect adherent host cells and change their morphology, causing the detachment of the morphed cells from the electrodes. Over time, as more cells get infected, the host cell-mediated cellular impedance decreases.
Plot the cell index, which is the change in impedance due to infected cell detachment, over time.
