Preparation of Mouse Bladder Tissue for Visualization of Bacterial Infection

Begin with a euthanized mouse with an exposed bladder, pre-infected with pathogenic bacteria.

The bacteria invade the urothelium and form colonies. Upon maturation, these bacteria rupture the epithelial cell and infect the surrounding cells.

Insert a catheter connected to a syringe filled with fixative. Slowly instill the fixative to inflate the bladder and preserve its architecture.

Clamp the urethra to retain the fixative and maintain bladder shape.

Remove the catheter, then cut the urethra. Transfer the bladder to fresh fixative and incubate to preserve the structure.

Section the bladder into two half-bladder cups to expose the urothelium, and rinse with a buffer.

Stain with a metal-based reagent to enhance contrast, then wash with ultrapure water.

Dehydrate the sections using an ethanol gradient, followed by critical-point drying.

Cut each dried bladder cup in half.

The sample is now ready for downstream processing to visualize host-pathogen interactions.

Once tuberculin slip-tip syringe with fixative, affix a catheter to the end, bevel facing opposite the syringe markings.

Snip off the excess tubing 1 to 2 millimeters from the end of the needle, taking care not to expose the needle tip.