Low Efficiency Upconversion Nanoparticles for High-Resolution Coalignment of Near-Infrared and Visible Light Paths on a Light Microscope.
Low Efficiency Upconversion Nanoparticles for High-Resolution Coalignment of Near-Infrared and Visible Light Paths on a Light Microscope.
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DOI：
10.1021/acsami.6b15322
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发表时间：
2017-03-08
影响因子：
9.5
通讯作者：
Canman JC
中科院分区：
文献类型：
--
作者：
Sundaramoorthy S;Garcia Badaracco A;Hirsch SM;Park JH;Davies T;Dumont J;Shirasu-Hiza M;Kummel AC;Canman JC
在光学显微镜中结合近红外(NIR)和可见光波长进行生物学研究越来越普遍。例如，生物学的许多领域都在开发将近红外用于光遗传学，即近红外激光诱导基因表达和/或蛋白质功能的变化。在光学显微镜上同时使用近红外和可见光的一个主要技术障碍是在成像平面位置获得它们的精确共对准。光子上转换粒子(UCP)在近红外光的激发下，通过反斯托克斯发光机制在可见光范围内发射，可以弥补这一缺陷。在这里，已经确定了两种不同的UCP，高效的micro540-UCP和低效率的Nan545-UCP，即使在非常高的近红外功率密度(>；25,000太阳)下，也能响应近红外光并发出具有高光稳定性的可见光。这两种UCP都可以被可见光和近红外光快速且可逆地激发，并发射可用绿色荧光蛋白(GFP)标准发射设置检测到的可见波长的光。绿色荧光蛋白是一种常用的基因编码荧光团。然而，高效率的micro540-UCP在近红外和可见光共对准方面并不理想，因为它们的尺寸更大，粒子到粒子之间的能量转移的空间展宽与长寿命激发态和饱和功率依赖一致。相比之下，效率较低的纳米UCP在近红外光束与可见光路径(分别为~2微米和~8微米光束展宽)的精确共对准方面具有优势，符合有限的粒子到粒子的能量转移，发射的超线性功率依赖，以及更小的粒子尺寸。此外，在酿酒酵母体内红外激光诱发基因操作符(IR-LEGO)的光遗传学分析中，纳米UCPs在近红外和可见光路径比对方面优于传统的双摄像头方法。总而言之，纳米UCP是一种强大的新工具，可以在光学显微镜上同时对准近红外和可见光路径。
The combination of near infrared (NIR) and visible wavelengths in light microscopy for biological studies is increasingly common. For example, many fields of biology are developing the use of NIR for optogenetics, in which an NIR laser induces a change in gene expression and/or protein function. One major technical barrier in working with both NIR and visible light on an optical microscope is obtaining their precise coalignment at the imaging plane position. Photon upconverting particles (UCPs) can bridge this gap as they are excited by NIR light but emit in the visible range via an anti-Stokes luminescence mechanism. Here, two different UCPs have been identified, high-efficiency micro540-UCPs and lower efficiency nano545-UCPs, that respond to NIR light and emit visible light with high photostability even at very high NIR power densities (>25,000 Suns). Both of these UCPs can be rapidly and reversibly excited by visible and NIR light and emit light at visible wavelengths detectable with standard emission settings used for Green Fluorescent Protein (GFP), a commonly used genetically-encoded fluorophore. However, the high efficiency micro540-UCPs were suboptimal for NIR and visible light coalignment, due to their larger size and spatial broadening from particle-to-particle energy transfer consistent with a long lived excited state and saturated power dependence. In contrast, the lower efficiency nano-UCPs were superior for precise coalignment of the NIR beam with the visible light path (~2 µm versus ~8 µm beam broadening respectively) consistent with limited particle-to-particle energy transfer, superlinear power dependence for emission, and much smaller particle size. Furthermore, the nano-UCPs were superior to a traditional two-camera method for NIR and visible light path alignment in an in vivo Infrared-Laser-Evoked Gene Operator (IR-LEGO) optogenetics assay in the budding yeast S. cerevisiae. In summary, nano-UCPs are powerful new tools for coaligning NIR and visible light paths on a light microscope.
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影响因子：
4.6
作者：
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通讯作者：
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影响因子：
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影响因子：
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作者：
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通讯作者：
Canman, Julie C.