Inflammation is a highly dynamic process with immune cells that continuously interact with each other and parenchymal components as they migrate through tissue. The dynamic cellular responses and interaction patterns are a function of the complex tissue environment that cannot be fully reconstructed ex vivo, making it necessary to assess cell dynamics and changing spatial patterning in vivo. These dynamics often play out deep within tissues, requiring the optical focus to be placed far below the surface of an opaque organ. With the emergence of commercially available two-photon excitation lasers that can be combined with existing imaging systems, new avenues for imaging deep tissues over long periods of time have become available. We discuss a selected subset of studies illustrating how two-photon microscopy (2PM) has helped to relate the dynamics of immune cells to their in situ function and to understand the molecular patterns that govern their behavior in vivo. We also review some key practical aspects of 2PM methods and point out issues that can confound the results, so that readers can better evaluate the reliability of conclusions drawn using this technology.

中文翻译：

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炎症研究中的动态多重组织成像


炎症是一个高度动态的过程，免疫细胞在组织中迁移时不断相互作用，并与实质成分相互作用。动态细胞反应和相互作用模式是复杂组织环境的功能，无法在体外完全重建，因此有必要评估体内细胞动力学和改变空间模式。这些动力学通常在组织深处发挥作用，需要将光学焦点放置在不透明器官表面的远下方。随着可与现有成像系统相结合的市售双光子激发激光器的出现，长期对深部组织进行成像的新途径已经出现。我们讨论了一组选定的研究，说明了双光子显微镜 （2PM） 如何帮助将免疫细胞的动力学与其原位功能联系起来，并了解控制其体内行为的分子模式。我们还回顾了 2PM 方法的一些关键实践方面，并指出了可能混淆结果的问题，以便读者可以更好地评估使用该技术得出的结论的可靠性。