Immunobiology and Therapy Unit, Institut Pasteur, INSERM U1224, Paris, France; Université Paris Cité, Paris, France.
Immunobiology and Therapy Unit, Institut Pasteur, INSERM U1224, Paris, France.
STAR Protoc. 2023 Mar 17;4(1):102062. doi: 10.1016/j.xpro.2023.102062. Epub 2023 Jan 24.
Mycobacterium tuberculosis (Mtb) hijacks host-derived fatty acids (FAs) to sustain its intracellular growth inside host cells. Here, we present a click-chemistry-based protocol to assess FA import by Mtb in axenic culture or inside mouse macrophages. We describe the use of alkyne analogs of natural FAs as an alternative to structurally altered fluorescent derivatives or hazardous radiolabeled FAs. We also detail quantitative analyses of FA uptake at single bacterial or host cell level by flow cytometry and confocal fluorescence microscopy. For complete details on the use and execution of this protocol, please refer to Laval et al. (2021)..
结核分枝杆菌（Mtb）劫持宿主来源的脂肪酸（FAs）以在宿主细胞内维持其细胞内生长。在这里，我们提出了一种基于点击化学的方案，以评估 Mtb 在体外培养或在小鼠巨噬细胞内的 FA 摄取。我们描述了使用炔烃类似物作为天然 FAs 的替代物，而不是结构改变的荧光衍生物或危险的放射性标记 FAs。我们还详细介绍了通过流式细胞术和共聚焦荧光显微镜对单个细菌或宿主细胞水平的 FA 摄取进行定量分析。有关此方案的使用和执行的完整详细信息，请参阅 Laval 等人。（2021）。
