Expanding Two-Photon Intravital Microscopy to the Infrared by Means of Optical Parametric Oscillator
Expanding Two-Photon Intravital Microscopy to the Infrared by Means of Optical Parametric Oscillator
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DOI：
10.1016/j.bpj.2009.10.035
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发表时间：
2010-02-17
影响因子：
3.4
通讯作者：
Niesner, Raluca A.
中科院分区：
文献类型：
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作者：
Herz, Josephine;Siffrin, Volker;Niesner, Raluca A.
大脑等各种器官中的慢性炎症意味着不同的免疫细胞亚群与靶器官的细胞相互作用为了从形态和功能上监测这种细胞通讯，可视化深层组织中两种以上颜色的能力是必不可少的在这里，我们证明了光学参量振荡器（OPO）的显著功率为基础的两个，光子激光扫描显微镜，用于中枢神经和免疫系统难以接近的器官的动态活体成像，特别适用于病理机制的长期研究（例如，慢性神经炎症）需要使用荧光蛋白将波长激发进一步扩展到红外克服了标准钛蓝宝石激光激发的当前限制，导致1），具有很大不同的激发和发射光谱的荧光团的同时成像（例如，GFP衍生物和RFP衍生物），以及2），组织中更高的穿透深度（高达80%），分辨率更高，光漂白和光毒性更低。组织成像，并将产生巨大的影响，蛋白质荧光团的选择，活体应用在生物科学和生物医学，因为我们在这项工作中证明
Chronic inflammation in various organs, such as the brain, implies that different subpopulations of immune cells interact with the cells of the target organ To monitor this cellular communication both morphologically and functionally, the ability to visualize more than two colors in deep tissue is indispensable Here, we demonstrate the pronounced power of optical parametric oscillator (OPO)-based two-photon laser scanning microscopy for dynamic intravital imaging in hardly accessible organs of the central nervous and of the immune system, with particular relevance for long-term investigations of pathological mechanisms (e g, chronic neuroinflammation) necessitating the use of fluorescent proteins Expanding the wavelength excitation farther to the infrared overcomes the current limitations of standard Titanium Sapphire laser excitation, leading to 1), simultaneous imaging of fluorophores with largely different excitation and emission spectra (e g, GFP-derivatives and RFP-derivatives), and 2), higher penetration depths in tissue (up to 80%) at higher resolution and with reduced photobleaching and phototoxicity This tool opens up new opportunities for deep-tissue imaging and will have a tremendous impact on the choice of protein fluorophores for intravital applications in bioscience and biomedicine, as we demonstrate in this work