Propagation and Harvest of Influenza Virus from a Fertilized Chicken Egg

Begin with an incubator containing a fertilized chicken egg.
The allantoic cavity, a fluid-filled compartment lined with epithelial cells rich in blood vessels, is pre-inoculated with influenza viruses.
During incubation, the virus interacts with the epithelial cells and is internalized via endocytosis.
Next, the virus releases viral RNA segments bound to nucleoproteins into the cytoplasm.
The released viral RNA acts as a template for new viral RNA and protein synthesis.
The synthesized viral RNAs and proteins assemble into new viruses, which bud into the allantoic fluid.
Incubate the egg at a low temperature to kill the embryo and constrict the blood vessels.
Place the egg with the air sac facing up and disinfect it.
Open the eggshell, and remove the protective membrane.
Move the embryo and yolk sac aside and collect the virus-rich fluid in a tube.
Centrifuge to separate debris, collect the virus-rich supernatant, and store it at a low temperature for further experiments.
Incubate the infected eggs at 37 degrees Celsius and 60% humidity without turning for the optimal incubation time, depending on the influenza virus type.
At the end of the incubation period, chill the eggs for a minimum of f