Measurement of 3-photon excitation and emission spectra and verification of Kasha's rule for selected fluorescent proteins excited at the 1700-nm window
Measurement of 3-photon excitation and emission spectra and verification of Kasha's rule for selected fluorescent proteins excited at the 1700-nm window
复制标题
测量 3 光子激发和发射光谱，并验证在 1700 nm 窗口激发的选定荧光蛋白的 Kasha 规则
DOI：
10.1364/oe.27.012723
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发表时间：
2019-04-29
期刊：
影响因子：
3.8
通讯作者：
Wang, Ke
中科院分区：
文献类型：
--
作者：
Deng, Xiangquan;Zhuang, Ziwei;Wang, Ke
荧光蛋白被广泛用于可视化各种生物样品中的结构和动力学。多光子显微镜特别适合于以亚细胞分辨率对表达荧光蛋白的结构进行成像。3-在1700-nm窗口激发的光子显微术（3 PM）已被证明对于表达红色荧光蛋白的深层组织（例如脑）成像是有希望的。然而，在这个窗口适合的荧光蛋白的3-光子激发和发射光谱仍然在很大程度上是未知的，阻碍了蛋白质的选择和检测优化。在这里，我们展示了3-光子激发和选定的荧光蛋白，适合在1700 nm窗口的3-光子激发发射光谱的详细测量。所测得的三光子激发光谱将为蛋白质和激发波长的选择提供指导。测量的三光子发射光谱以及与单光子发射光谱的比较，一方面证明了基本的Kasha规则对这些荧光蛋白的三光子荧光仍然有效，另一方面将有助于有效的荧光信号检测。(C)根据OSA开放获取出版协议的条款，2019年美国光学学会
Fluorescent proteins are widely used to visualize structures and dynamics in various biological samples. Multiphoton microscopy is especially suitable for imaging structures expressing fluorescent proteins with subcellular resolution. 3-photon microscopy (3PM) excited at the 1700-nm window has proven to be promising for deep-tissue (such as brain) imaging expressing red fluorescent proteins. However, the 3-photon excitation and emission spectra of fluorescent proteins suitable at this window remain largely unknown, hampering protein selection and detection optimization. Here we demonstrate detailed measurement of 3-photon excitation and emission spectra for selected fluorescent proteins, suitable for 3-photon excitation at the 1700-nm window. The measured 3-photon excitation spectra will provide guidelines for protein and excitation wavelength selection. The measured 3-photon emission spectra and comparison with the 1-photon emission spectra, on one hand proves that the fundamental Kasha's rule is still valid for 3-photon fluorescence in these fluorescent proteins, on the other hand will be helpful for efficient fluorescence signal detection. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement