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Microbiology JoVE (Open Access) Citable · DOI

4D Multimodality Imaging of Citrobacter rodentium Infections in Mice

DOI: 10.3791/50450-v
What you'll learn
  • ✓ Prepare bioluminescent Citrobacter rodentium inoculum for mouse infection
  • ✓ Perform oral gavage infection and monitor bacterial colonization longitudinally
  • ✓ Acquire and reconstruct 4D multimodality imaging data (DLIT-μCT)
  • ✓ Generate 4D spatiotemporal visualizations of infection dynamics
Protocol

Biopharma Insights Multi-modality imaging is a valuable approach for studying bacterial colonization in small animal models. This protocol outlines infection of mice with bioluminescent Citrobacter rodentium and the longitudinal monitoring of bacterial colonization using composite 3D diffuse light imaging tomography with μCT imaging to create a 4D movie of C. rodentium infection.

Difficulty
advanced
Total time
~10–14 days (infection + daily imaging for 5–7 days)
Model organism
Mouse (strain not specified; C. rodentium susceptible strain recommended)
Biosafety
BSL-2

Steps

1
Prepare bioluminescent Citrobacter rodentium inoculum

Culture and prepare bioluminescent C. rodentium to appropriate density for infection. Verify bacterial viability and luminescence output before mouse inoculation.

▶ 01:21
2
Infect mice with bioluminescent C. rodentium

Administer bioluminescent C. rodentium to mice via oral gavage or appropriate route. Record infection parameters (dose, strain, cfu) for reproducibility.

▶ 02:44
3
Perform daily DLIT-μCT multimodality imaging

Acquire composite 3D diffuse light imaging tomography (DLIT) and micro-computed tomography (μCT) scans daily to longitudinally monitor bacterial colonization in live mice.

▶ 04:09
4
Reconstruct 3D DLIT-μCT imaging data

Process and align raw DLIT and μCT image datasets using appropriate reconstruction software to generate co-registered 3D volumetric images.

▶ 06:15
5
Generate 4D movie of infection dynamics

Compile reconstructed daily 3D datasets into a temporal 4D visualization to spatially and temporally track C. rodentium colonization throughout infection.

▶ 09:29
6
Analyze monitored infection with defined inoculum

Present representative imaging results and infection progression data from mice inoculated with 5 × 10⁹ cfu C. rodentium strain ICC180 to demonstrate protocol feasibility.

▶ 10:24
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