Simultaneous multimodal three-photon and optical coherence microscopy of the mouse brain in the 1700 nm optical window in vivo.
Simultaneous multimodal three-photon and optical coherence microscopy of the mouse brain in the 1700 nm optical window in vivo.
复制标题
在体内 1700 nm 光学窗口中同时进行多模态三光子和光学相干显微镜观察小鼠大脑。
DOI：
10.1101/2023.09.11.557176
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Xu,Chris
中科院分区：
文献类型：
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作者：
Yang,Xusan;Liu,Siyang;Xia,Fei;Wu,Meiqi;Adie,Steven;Xu,Chris
多模态显微镜结合各种成像方法可以在单个成像会话中提供关于组织的互补信息。在这里，我们展示了一种多模式的方法相结合的三光子显微镜（3 PM）和光谱域光学相干显微镜（SD-OCM）。我们表明，光参量啁啾脉冲放大（OPCPA）激光源，这是三光子荧光激发和三次谐波产生（THG）的标准源，可用于同时OCM，3光子（3 P）荧光和THG成像。我们验证了系统的性能，在小鼠脑深部在体内与OPCPA源工作在1620 nm的中心波长。我们在完整的成年小鼠脑中使用线性和非线性对比度以高空间分辨率非侵入性地在>1 mm深度处可视化白色物质中的小结构，例如有髓轴突、神经元和大纤维束。我们的研究结果表明，同时在长波长窗口的OCM和3 PM可以方便地结合在体内深部组织成像。
Multimodal microscopy combining various imaging approaches can provide complementary information about tissue in a single imaging session. Here, we demonstrate a multimodal approach combining three-photon microscopy (3PM) and spectral-domain optical coherence microscopy (SD-OCM). We show that an optical parametric chirped-pulse amplification (OPCPA) laser source, which is the standard source for three-photon fluorescence excitation and third harmonic generation (THG), can be used for simultaneous OCM, 3-photon (3P) fluorescence and THG imaging. We validated the system performance in deep mouse brains in vivo with an OPCPA source operating at 1620 nm center wavelength. We visualized small structures such as myelinated axons, neurons, and large fiber tracts in white matter with high spatial resolution non-invasively using linear and nonlinear contrast at >1 mm depth in intact adult mouse brain. Our results showed that simultaneous OCM and 3PM at the long wavelength window can be conveniently combined for deep tissue imaging in vivo.