JoVE Encyclopedia of Experiments
Microbiology
0 views • 6:10 min • October 30th, 2025
Take a multi-well plate containing sterile human urine.
Inoculate the test wells with bacteria. The control wells contain urine alone.
Take a 3D-printed, silicone-coated device mimicking a catheter’s inner surface, where biofilms develop during catheter-associated urinary infections.
Place it into the wells, seal the plate, and incubate it.
Bacteria attach to the device surface, utilize nutrients from the urine, and form biofilms.
Rinse the device with a buffer to remove loosely bound bacteria.
Place the device into a multi-well plate containing crystal violet, which interacts with the biofilm to stain it.
Rinse with water and dry it.
Immerse the device in a multi-well plate containing an acid solution to disrupt dye-biofilm interactions and release the dye into the solution.
Measure the absorbance of the solutions in each well using a spectrophotometer.
Higher absorbance in the test wells compared to the control wells confirms biofilm formation on the device, indicating its suitability to model catheter-associated urinary tract infection.
To begin, obtain pooled human urine sample. Aliquot 135 microliters of pooled human urine into each on-test well of a 96-well plate following the designate
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