Intravital two-photon imaging: a versatile tool for dissecting the immune system
Intravital two-photon imaging: a versatile tool for dissecting the immune system
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DOI：
10.1136/ard.2010.138156
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发表时间：
2011-03-01
影响因子：
27.4
通讯作者：
Ishii, Masaru
中科院分区：
文献类型：
--
作者：
Ishii, Taeko;Ishii, Masaru
在过去的十年中，多光子或“双光子”激发显微镜开启了生物成像领域的新时代。用于双光子显微镜的近红外激发激光可以穿透较厚的标本，无需进行薄切片即可可视化组织和器官深处的活细胞行为。最大限度地减少光漂白和毒性，使活体和完整标本能够长时间可视化。在这篇简短的综述中，讨论了免疫系统生理学和病理学的活体双光子成像的最新发现。免疫系统配置了高度动态的网络，其中许多细胞类型活跃地在全身移动并在特定区域相互作用。因此，实时活体成像可能是剖析该动态系统机制的有力工具。免疫系统最独特的特征是其高度动态的性质。多种细胞类型，例如淋巴细胞、巨噬细胞和树突状细胞（DC）在全身不断循环，通过外周组织迁移并在各自的生态位中相互作用。尽管整个动物中免疫细胞的动态仍不太清楚，但传统的免疫学方法，如流式细胞术、细胞或组织培养、生物化学和组织学，已在该领域取得了巨大的成就。荧光显微镜技术的进步使我们能够可视化尚未研究的完整生物现象。在这些进步中，最近出现和流行的双光子激发激光显微镜彻底改变了研究领域，使得活体器官深处细胞的动态行为可以被可视化和分析。
During the past decade, multi-photon or 'two-photon' excitation microscopy has launched a new era in the field of biological imaging. The near-infrared excitation laser for two-photon microscopy can penetrate thicker specimens, enabling the visualisation of living cell behaviour deep within tissues and organs without thin sectioning. The minimised photobleaching and toxicity enables the visualisation of live and intact specimens for extended periods. In this brief review, recent findings in intravital two-photon imaging for the physiology and pathology of the immune system are discussed. The immune system configures highly dynamic networks, where many cell types actively travel throughout the body and interact with each other in specific areas. Hence, real-time intravital imaging may be a powerful tool for dissecting the mechanisms of this dynamic system.The most unique characteristic of the immune system is its highly dynamic nature. A variety of cell types, such as lymphocytes, macrophages and dendritic cells (DCs), are continuously circulating throughout the body, migrating through the peripheral tissues and interacting with each other in their respective niches. Conventional methodologies in immunology, such as flow cytometry, cell or tissue culture, biochemistry and histology, have brought tremendous achievement within this field, although the dynamics of immune cells in an entire animal remain less clear.Technological progress of fluorescence microscopy has enabled us to visualise the intact biological phenomenon that has been uninvestigated. Among the advancements, the recent emergence and prevalence of two-photon, excitation-based, laser microscopy has revolutionised the research field, such that the dynamic behaviour of cells deep inside living organs can be visualised and analysed.