title: 4D multimodality imaging of Citrobacter rodentium infections in mice.
abstract: This protocol outlines the steps required to longitudinally monitor a bioluminescent bacterial infection using composite 3D diffuse light imaging tomography with integrated μCT (DLIT-μCT) and the subsequent use of this data to generate a four dimensional (4D) movie of the infection cycle. To develop the 4D infection movies and to validate the DLIT-μCT imaging for bacterial infection studies using an IVIS Spectrum CT, we used infection with bioluminescent C. rodentium, which causes self-limiting colitis in mice. In this protocol, we outline the infection of mice with bioluminescent C. rodentium and non-invasive monitoring of colonization by daily DLIT-μCT imaging and bacterial enumeration from feces for 8 days. The use of the IVIS Spectrum CT facilitates seamless co-registration of optical and μCT scans using a single imaging platform. The low dose μCT modality enables the imaging of mice at multiple time points during infection, providing detailed anatomical localization of bioluminescent bacterial foci in 3D without causing artifacts from the cumulative radiation. Importantly, the 4D movies of infected mice provide a powerful analytical tool to monitor bacterial colonization dynamics in vivo.
authors: MRC Centre for Molecular Bacteriology and Infection, Division of Cell & Molecular Biology, Imperial College London.
authors: James William
authors: JW
authors: Collins
authors: Jeffrey A
authors: JA
authors: Meganck
authors: Chaincy
authors: C
authors: Kuo
authors: Kevin P
authors: KP
authors: Francis
authors: Gad
authors: G
authors: Frankel
mesh_terms: D000818:Animals; D044082:Citrobacter rodentium / Q000737:chemistry / Q000254:growth & development / Q000302:isolation & purification*; D004756:Enterobacteriaceae Infections / Q000175:diagnosis / Q000382:microbiology*; D021621:Imaging, Three-Dimensional / Q000379:methods; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D064847:Multimodal Imaging / Q000379:methods*; D041622:Tomography, Optical / Q000379:methods*
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