Visualizing Intracellular Bacterial Communities in an Infected Mouse Bladder

JoVE Encyclopedia of Experiments

Microbiologie

0 weergaven • 2:06 min • March 31st, 2026

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

Start with an immobilized, dissected mouse bladder pre-infected with β-galactosidase-expressing uropathogenic E. coli, placed in a silicone-coated multi-well plate.

The bladder’s lumen faces up, exposing the epithelial cells that contain intracellular bacterial communities, or IBCs.

Wash the bladder with a buffer to remove debris and unattached bacteria.

Next, add a fixative and incubate to preserve cellular and bacterial structures.

Remove the fixative and wash with a buffer.

Then, wash with a detergent-containing buffer to permeabilize cellular and bacterial membranes.

Remove the buffer, then add a solution of X-gal, a substrate of β-galactosidase, supplemented with redox reagents, and incubate.

X-gal and redox reagents enter the bacteria. β-galactosidase cleaves X-gal, producing a colorless intermediate that is oxidised by redox reagents to form a blue precipitate.

Under a dissecting microscope, the blue dots in the bladder confirm the IBCs, indicating a urinary tract infection.

For IBC enumeration, after harvesting the bladder, place it in a silicone-coated well of a 6-well plate with PBS and use scissors to cut it in half.

Using small metal pins placed along the outermost edges, gently splay the bladder so that the lumen is facing upward and the maximum amount of urothelium is exposed. After performing LacZ stain as outlined in the text protocol, observe IBCs, which appear as blue puncta under a dissecting microscope.

10:23

Urineweginfectie in een kleine diermodel: transuretrale catheterisatie van mannelijke en vrouwelijke muizen

Gerelateerde video's

0 Bekeken

06:09

Intrarenale injectie van Escherichia coli In een Rat Model van Pyelonefritis

Gerelateerde video's

0 Bekeken

07:34

Isolatie van één intracellulaire bacteriële gemeenschappen gegenereerd op basis van een lymfkliertest Model van urine landstreekbesmetting Downstream eencellige p.a.

Gerelateerde video's

0 Bekeken

09:24

Transurethrale Inductie van Mouse urineweginfectie

Gerelateerde video's

0 Bekeken

02:56

Isolation of Mouse Bladder Epithelial Cells Containing Intracellular Bacterial Communities

Gerelateerde video's

0 Bekeken

03:14

Visualization of Urinary Tract Infection in a Mouse Model Using Bioluminescence

Gerelateerde video's

0 Bekeken

05:23

Preparation of Mouse Bladder Tissue for Visualization of Bacterial Infection

Gerelateerde video's

0 Bekeken

02:32

Transurethral Catheterization and Bacterial Inoculation to Generate a Urinary Tract Infection Model in Male Mice

Gerelateerde video's

0 Bekeken

03:43

Quantifying Immune Cell Infiltration in Uropathogenic E. coli-Infected Mouse Bladder Tissue using Flow Cytometry

Gerelateerde video's

0 Bekeken

06:10

Visualization of Bacteria in Bladder Biopsy Sections via Fluorescence In Situ Hybridization

Gerelateerde video's

0 Bekeken

Laatst bijgewerkt: 15 augustus 2026