JoVE Encyclopedia of Experiments
Microbiology
0 views • 3:41 min • August 31st, 2026
Begin with tubes containing human lung epithelial cells.
Add biotinylated influenza viruses and incubate on ice. The viruses bind to the cell surface without being internalized.
Centrifuge to separate the virus-bound cells, remove the supernatant and resuspend.
In the initial time point tube, add unlabeled streptavidin, which binds to the biotinylated viruses, forming complexes.
Add fluorophore-labeled streptavidin, which fails to bind to the streptavidin-biotin virus complex. Wash and fix the cells.
Incubate the later time-point tube at a higher temperature, enabling virus internalization, then return to ice.
Add unlabeled streptavidin and an internalization-blocking agent, which prevents further viral uptake and incubate. Streptavidin binds surface-bound biotinylated viruses, then wash.
Fix the cells and add a detergent-containing buffer to permeabilize them.
Wash and add fluorophore-labeled streptavidin, which enters the cells and binds to biotin.
Analyze the samples by flow cytometry.
A higher fluorescence in the later time point confirms successful viral internalization.
After repeating the wash one more time, cool the tubes down on ice for 10 minutes. When the tubes have cooled, spin ag
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