Germain Ronald N, Castellino Flora, Chieppa Marcello, Egen Jackson G, Huang Alex Y C, Koo Lily Y, Qi Hai
Lymphocyte Biology Section, Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, DHHS, Bldg. 10 Rm. 11N311, 10 Center Dr. MSC-1892 Bethesda, MD 20892, USA.
Semin Immunol. 2005 Dec;17(6):431-41. doi: 10.1016/j.smim.2005.09.003. Epub 2005 Oct 10.
The past few years have seen the application of confocal and especially two-photon microscopy to the dynamic high-resolution imaging of lymphocytes and antigen presenting cells within organs such as lymph nodes and thymus. After summarizing some of the published results obtained to date using these methods, we describe our view of how this technology will develop and be applied in the near future. This includes its extension to a wide variety of non-lymphoid tissues, to the tracking of functional responses in addition to migratory behavior, to the analysis of molecular events previously studied only in vitro, to dissection of the interplay between hematopoietic and stromal elements, to visualization of a wider array of cell types including neutrophils, macrophages, NK cells, NKT cells and others, and to the interaction of the host with infectious agents. Reaching these goals will depend on a combination of new tools for genetic manipulations, novel fluorescent reporters, enhanced instrumentation, and better surgical techniques for the extended imaging of live animals. The end result will be a new level of understanding of how orchestrated cell movement and interaction contribute to the physiological and pathological activities of the immune system.
在过去几年中,共聚焦显微镜,尤其是双光子显微镜,已被应用于对淋巴结和胸腺等器官内淋巴细胞和抗原呈递细胞进行动态高分辨率成像。在总结了迄今为止使用这些方法所获得的一些已发表结果之后,我们阐述了我们对这项技术在不久的将来如何发展和应用的看法。这包括将其扩展到多种非淋巴组织,除了迁移行为之外,还用于追踪功能反应,用于分析以前仅在体外研究的分子事件,用于剖析造血和基质成分之间的相互作用,用于可视化更广泛的细胞类型,包括中性粒细胞、巨噬细胞、自然杀伤细胞、自然杀伤T细胞等,以及用于宿主与感染因子之间的相互作用。要实现这些目标,将取决于基因操作的新工具、新型荧光报告分子、增强的仪器设备以及用于活体动物扩展成像的更好手术技术的综合运用。最终结果将是对精心编排的细胞运动和相互作用如何促进免疫系统的生理和病理活动有一个全新的认识水平。