Facultative intracellular pathogens are able to live inside and outside host cells. It is highly desirable to differentiate their cellular locations for the purposes of fundamental research and clinical applications. In this work, we developed a novel analysis platform that allows users to choose two analysis models: amplitude weighted lifetime ( τ A ) and intensity weighted lifetime ( τ I ) for fluorescence lifetime imaging microscopy (FLIM). We applied these two models to analyse FLIM images of mouse Raw macrophage cells that were infected with bacteria Shigella Sonnei , adherent and invasive E. coli (AIEC) and Lactobacillus . The results show that the fluorescence lifetimes of bacteria depend on their cellular locations. The τ A model is superior in visually differentiating bacteria that are in extra- and intra-cellular and membrane-bounded locations, whereas the τ I model show excellent pr...

中文翻译：

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使用双光子荧光寿命成像显微镜研究细菌细胞位置的快速分析平台

兼性细胞内病原体能够生活在宿主细胞的内部和外部。为了基础研究和临床应用的目的，非常需要区分它们的细胞位置。在这项工作中，我们开发了一个新颖的分析平台，允许用户选择两种分析模型：用于荧光寿命成像显微镜（FLIM）的振幅加权寿命（τA）和强度加权寿命（τI）。我们应用了这两个模型来分析小鼠原始巨噬细胞的FLIM图像，这些细胞感染了细菌志贺氏菌，黏附性和侵袭性大肠杆菌（AIEC）和乳杆菌。结果表明，细菌的荧光寿命取决于其细胞位置。