JoVE Encyclopedia of Experiments
Microbiology
0 views • 3:47 min • February 26th, 2026
Begin with two glass-bottom microdishes containing suspensions of a biofilm-forming bacterium.
The treatment dish contains an active signaling-disrupting enzyme, while the control includes the inactivated enzyme.
Cover the dishes and incubate to allow bacterial adhesion to the microdish surface.
Wash to remove non-adherent cells, then introduce fresh medium containing the enzymes and incubate.
In the control dish, the bacteria communicate through signaling molecules and produce extracellular polymeric substances (EPS), forming a biofilm.
The active enzyme degrades the signaling molecules, disrupting their function and reducing biofilm formation.
Replenish with fresh medium and incubate further to reinforce the enzyme's biofilm-disrupting effects.
Remove the medium, add fluorescently labeled lectins, and incubate to allow lectin binding to the EPS.
Wash to remove any unbound lectin.
Incubate with a fixative to preserve biofilm architecture, then rinse the dish.
Acquire confocal microscopy images to assess the enzyme-mediated suppression of biofilm formation.
To carry out confocal laser scanning microscopy, or CLSM, of A. baumannii S1 biofilms, grow a 5 milliliter culture of A. baumannii S1 in
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