Bacterial infections associated with implanted medical devices represents a healthcare crisis due to their persistence, antibiotic tolerance, and immune avoidance. Indwelling devices are rapidly coated with host plasma and extracellular matrix proteins which can then be exploited by bacterial pathogens for adherence and subsequent biofilm development. Our understanding of the host-pathogen interface that determines the fate of biofilm-mediated infections is limited to the experimental models employed by laboratories studying these organisms. Current in vivo models of biofilm-mediated infection, while certainly useful, are typically limited to end-point analyses of bacterial burden enumeration, immune cell profiling, and cytokine/chemokine analysis. Thus, with these models, the complex, real-time assessment of biofilm development and innate immune cell activity remains imperceptible. Here, we describe a novel murine biofilm infection model employing time-lapse intravital multiphoton microscopy which permits concurrent and real-time visualization of Staphylococcus aureus biofilm formation and immune cell activity. Using cell tracking, we found that S. aureus biofilms impede neutrophil chemotaxis, redirecting their migration patterns to prevent biofilm invasion. This approach is the first to directly examine device-associated biofilm development and host-pathogen interactions and will serve to both further our understanding of infection development and help reveal the effects of future antibiofilm treatment strategies.

与植入的医疗设备相关的细菌感染由于其持久性，抗生素耐受性和免疫避免性，代表了医疗保健危机。留置装置被宿主血浆和细胞外基质蛋白迅速包被，然后细菌病原体可以利用它们粘附和随后形成生物膜。我们对决定生物膜介导的感染命运的宿主-病原体界面的了解仅限于研究这些生物的实验室采用的实验模型。当前体内生物膜介导的感染模型虽然肯定有用，但通常仅限于细菌负荷计数，免疫细胞谱分析和细胞因子/趋化因子分析的终点分析。因此，使用这些模型，对生物膜发育和先天免疫细胞活性的复杂，实时评估仍然难以察觉。在这里，我们描述了一种新型的小鼠生物膜感染模型，该模型采用延时活体多光子显微镜技术，可以同时并实时地观察金黄色葡萄球菌生物膜形成和免疫细胞活性。使用单元跟踪，我们发现金黄色葡萄球菌生物膜阻碍中性粒细胞趋化性，改变其迁移模式以防止生物膜入侵。这种方法是第一个直接检查与设备相关的生物膜发展和宿主-病原体相互作用的方法，将有助于我们进一步了解感染的发展，并有助于揭示未来抗生物膜治疗策略的效果。