In vivo deep-brain imaging of microglia enabled by three-photon fluorescence microscopy
In vivo deep-brain imaging of microglia enabled by three-photon fluorescence microscopy
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三光子荧光显微镜实现小胶质细胞体内深部脑成像
DOI：
10.1364/ol.408329
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发表时间：
2020-09-15
期刊：
影响因子：
3.6
通讯作者：
Wang, Ke
中科院分区：
文献类型：
--
作者：
Cheng, Hui;Tong, Shen;Wang, Ke
小胶质细胞是脑内主动免疫防御的第一和主要形式。然而，在动物模型中，由于缺乏深层成像技术，对这些细胞的研究仅限于大脑的表层。在这里，我们打破了这个深度限制，使用三光子荧光（3PF）显微镜激发在1700 nm的窗口。三光子作用截面（eta sigma（3））测量为染料选择和3PF产生的最大化奠定了基础。3PF成像抑制了表面背景，与常用的双光子显微镜（2PM）相比，大大提高了信号与背景比。我们可以在体内脑表面以下1124 μ m处对小胶质细胞进行成像，比以前使用2PM进行小胶质细胞成像的结果深3.7倍。这项技术使我们能够可视化小胶质细胞在体内的白色物质层的第一次。(C)2020美国光学学会
Microglia act as the first and main form of active immune defense in brain. However, in animal models, research on these cells is limited to the superficial layer of the brain, due to the lack of a deep-imaging technique. Here we break this depth limit using three-photon fluorescence (3PF) microscopy excited at the 1700-nm window. Three-photon action cross-section (eta sigma(3)) measurement lays the basis for dye selection and the resultant maximization of 3PF generation. 3PF imaging suppresses the surface background, leading to a much improved signal-to-background ratio compared to the commonly used two-photon microscopy (2PM). We can image microglia 1124 mu m below the brain surface in vivo, 3.7 times deeper than previous results using 2PM for microglia imaging. This technique enables us to visualize microglia in the white matter layer in vivo for the first time. (C) 2020 Optical Society of America