Department of Medicine 3-Rheumatology and Immunology, Friedrich-Alexander-University Erlangen-Nürnberg and Universitätsklinikum Erlangen, Erlangen, Germany.
Deutsches Zentrum für Immuntherapie, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany.
Annu Rev Pathol. 2024 Jan 24;19:43-67. doi: 10.1146/annurev-pathmechdis-070323-124158. Epub 2023 Sep 18.
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.
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。动态的细胞反应和相互作用模式是复杂组织环境的功能，这种环境无法在体外完全重建，因此需要在体内评估细胞动态和不断变化的空间模式。这些动态通常在组织深处发生，需要将光学焦点放置在不透明器官的表面以下很远的地方。随着可商用的双光子激发激光器的出现，这些激光器可以与现有的成像系统相结合，为长时间内对深层组织进行成像开辟了新途径。我们讨论了一组精选的研究，这些研究说明了双光子显微镜（2PM）如何帮助将免疫细胞的动态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，以便读者能够更好地评估使用该技术得出的结论的可靠性。
