An In Vivo Model to Study Shear-Dependent Bacterial Adhesion

Take an anesthetized mouse fasted overnight to reduce bowel movements.

Apply eye ointment to prevent corneal drying. Secure it on a heating pad to regulate body temperature.

Expose the right jugular vein. Insert a catheter and secure it with stitches.

Open the peritoneal cavity, then place the mouse on a transparent plate.

Spread the intestine to expose the mesenterium, a supportive membrane, to access the blood vessels.

Maintain body temperature and apply saline to prevent dehydration.

Place the mouse under a fluorescence microscope, then immobilize the vessels.

Apply an ionophore that elevates calcium levels in endothelial cells. This triggers the release of von Willebrand factor (VWF), forming coiled strands on the cell surface.

Inject fluorescently labeled bacteria through the catheter to reach the blood vessel.

The shear stress from blood flow uncoils the VWF strands, which bind to bacterial adhesins and form bacterial aggregates.

Capture time-lapse images to quantify shear-dependent bacterial adhesion.

Fast six to eight-week-old mice the night before the experiment to reduce bowel movement. The day of the experiment begin by injecting 0.1 milligrams of buprenorphine per kilogram of body