JoVE Encyclopedia of Experiments
Microbiology
0 views • 1:20 min • February 26th, 2026
Take cultures of human urinary bladder epithelial cells in a multiwell plate.
Incubate the control culture with wild-type Enterobacter cloacae, a pathogenic bacterium with pili, hair-like structures on the bacterial surface.
The FimH subunit of the pili binds to the host cell receptor, anchoring the wild-type bacteria to the host cell.
Incubate the experimental culture with a mutant Enterobacter cloacae lacking FimH.
Without FimH, the mutant bacteria fail to adhere to the host cells.
Post-incubation, wash with a buffer to remove the unbound bacteria.
Add a lysis buffer and incubate under agitation to disrupt the host cells and release the adhered bacteria.
Serially dilute the lysates, plate them on nutrient media, and incubate.
The lysate from the control culture yields colonies, while the lysate from the experimental culture does not, confirming that FimH is essential for bacterial adhesion to the host cells.