Snyder Institute for Chronic Diseases, University of Calgary, Calgary, AB, Canada.
Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada.
Methods Mol Biol. 2022;2440:165-180. doi: 10.1007/978-1-0716-2051-9_10.
The ability to visualize biological phenomenon has driven scientific interest and advancement over the centuries. Although many methods and assays provide a detailed snapshot of a physiology, the ability to track such processes in real time has expanded the breadth of questions that can be interrogated in the laboratory. Intravital Microscopy (IVM) is a dynamic and powerful way to investigate both the homeostatic and host response to either therapeutic or pathological intervention using live animals. In this technique, animal models, (often mice) are anesthetized, and the organ of interest surgically exteriorized. The animal containing fluorescent labels (either endogenous, or conjugated to antibodies/proteins) will then be placed on a high-powered laser scanning microscope, where the labeled cells or structures can be observed in their natural environment. Complex behavioral data and interactions can be captured in a temporal manner, providing a plethora of information that will help researchers make conclusions on a more systemic level, rather than isolating only part the response. As the technology advances, a greater number of imaging modality options can be utilized, and more diverse research questions can be addressed. The goal of this chapter is to highlight IVM as a technique and help instruct new users on how to choose the proper modalities, and by using imaging of a skin wound in mice as a model, provide troubleshooting strategies, technical advice, and considerations.
几个世纪以来，对生物现象进行可视化的能力推动了科学的发展和进步。尽管许多方法和检测手段可以详细地了解生理学，但实时跟踪这些过程的能力扩大了可以在实验室中研究的问题的范围。活体显微镜检查（IVM）是一种动态而强大的方法，可以使用活体动物研究生理稳态和对治疗或病理干预的宿主反应。在该技术中，动物模型（通常是小鼠）被麻醉，并通过手术将感兴趣的器官暴露出来。含有荧光标记物（内源性的或与抗体/蛋白质偶联的）的动物将被放置在高功率激光扫描显微镜上，在那里可以在其自然环境中观察到标记的细胞或结构。可以以时间方式捕获复杂的行为数据和相互作用，提供大量信息，帮助研究人员在更系统的层面上得出结论，而不是仅孤立地研究部分反应。随着技术的进步，可以利用更多的成像模式选择，并且可以解决更多的研究问题。本章的目的是强调 IVM 作为一种技术，并帮助新用户选择合适的模式，并通过使用小鼠皮肤伤口的成像作为模型，提供故障排除策略、技术建议和注意事项。
