Collins James William, Meganck Jeffrey A, Kuo Chaincy, Francis Kevin P, Frankel Gad
MRC Centre for Molecular Bacteriology and Infection, Division of Cell & Molecular Biology, Imperial College London.
J Vis Exp. 2013 Aug 13(78):50450. doi: 10.3791/50450.
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.
本方案概述了使用集成μCT的复合3D漫射光成像断层扫描（DLIT-μCT）纵向监测生物发光细菌感染所需的步骤，以及随后利用这些数据生成感染周期的四维（4D）电影的过程。为了制作4D感染电影并验证使用IVIS Spectrum CT进行细菌感染研究的DLIT-μCT成像，我们使用了生物发光的啮齿柠檬酸杆菌感染，该菌可在小鼠中引起自限性结肠炎。在本方案中，我们概述了用生物发光的啮齿柠檬酸杆菌感染小鼠，并通过每日DLIT-μCT成像和粪便细菌计数对定植进行8天的非侵入性监测。IVIS Spectrum CT的使用有助于在单个成像平台上实现光学和μCT扫描的无缝共配准。低剂量μCT模式能够在感染期间的多个时间点对小鼠进行成像，在不产生累积辐射伪影的情况下，提供生物发光细菌病灶在3D中的详细解剖定位。重要的是，感染小鼠的4D电影为监测体内细菌定植动态提供了一个强大的分析工具。
