Measure influenza viral titration with mKate2 fluorescent protein reporter in 96-well plate

  1. MDCK host cells were seeded in a 96-well microplate and allowed to adhere overnight
  2. A titration of 5-folds of mKate2-influenza viral particles were prepared and added to the host cells
  3. The samples were allowed to incubate for 24 hours and then imaged and analyzed with Celigo image cytometer to count the number of infected cells
  4. The Celigo software was used to identify the total MDCK cells in bright field images, and then using the fluorescent gating function to determine the percent infection rate
influenza viral titration

Bright field and fluorescent images of the individual infected cells in the 96-well.  The Celigo software gating shows the counting results of mKate2 positive infected cells.

influenza infectivity curve

Utilizing the gating functions in Celigo, the % mKate2 positive cells can be measured to generate an infectivity curve

The Celigo Image Cytometer automates viral titer assays: