Grupo de Microbiología Integrativa, Laboratorio de Biología Estructural y Molecular BEM, Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile.
Millenium Institute Center for Genome Regulation, Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile.
Methods Mol Biol. 2025;2852:171-179. doi: 10.1007/978-1-0716-4100-2_12.
Studying host-pathogen interactions is essential for understanding infectious diseases and developing possible treatments, especially for priority pathogens with increased virulence and antibiotic resistance, such as Klebsiella pneumoniae. Over time, this subject has been approached from different perspectives, often using mammal host models and invasive endpoint measurements (e.g., sacrifice and organ extraction). However, taking advantage of technological advances, it is now possible to follow the infective process by noninvasive visualization in real time, using optically amenable surrogate hosts. In this line, this chapter describes a live-cell imaging approach to monitor the interaction of K. pneumoniae and potentially other bacterial pathogens with zebrafish larvae in vivo. This methodology is based on the microinjection of fluorescent bacteria into the otic vesicle, followed by time-lapse observation by automated fluorescence microscopy with environmental control, monitoring the dynamics of immune cell recruitment, bacterial load, and larvae survival.
研究宿主-病原体相互作用对于理解传染病和开发可能的治疗方法至关重要，特别是对于具有毒力增加和抗生素耐药性的优先病原体，如肺炎克雷伯菌。随着时间的推移，这个主题已经从不同的角度进行了研究，通常使用哺乳动物宿主模型和侵入性终点测量（例如，牺牲和器官提取）。然而，利用技术进步，现在可以通过非侵入性实时可视化利用光学上可接受的替代宿主来跟踪感染过程。在这方面，本章描述了一种活细胞成像方法，用于监测肺炎克雷伯菌和潜在其他细菌病原体与体内斑马鱼幼虫的相互作用。该方法基于将荧光细菌微注射到耳囊中，然后通过自动荧光显微镜进行延时观察，并进行环境控制，监测免疫细胞募集、细菌负荷和幼虫存活的动态变化。
