Real-Time, Two-Color Stimulated Raman Scattering Imaging of Mouse Brain for Tissue Diagnosis.
Real-Time, Two-Color Stimulated Raman Scattering Imaging of Mouse Brain for Tissue Diagnosis.
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DOI：
10.3791/63484
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发表时间：
2022-02-01
期刊：
影响因子：
--
通讯作者：
Fu D
中科院分区：
文献类型：
--
作者：
Espinoza R;Wong B;Fu D
受激拉曼散射（SRS）显微镜已经成为用于组织诊断的强大光学成像技术。近年来，双色SRS已被证明能够提供苏木精和伊红（H&E）等效的图像，从而可以快速可靠地诊断脑癌。这样的能力已经实现了令人兴奋的术中癌症诊断应用。组织的双色SRS成像可以用皮秒或飞秒激光源完成。飞秒激光器的优点是能够实现灵活的成像模式，包括快速高光谱成像和实时双色SRS成像。具有啁啾激光脉冲的光谱聚焦方法通常与飞秒激光器一起使用以实现高光谱分辨率。利用正交调制和锁相检测技术可以实现双色SRS信号的采集。脉冲啁啾、调制和表征的复杂性是这种方法广泛采用的瓶颈。本文提供了一个详细的协议，以证明实施和优化的光谱聚焦SRS和实时，双色成像的小鼠脑组织在epi模式。该协议可用于利用SRS的高速和光谱成像能力的广泛的SRS成像应用。
Stimulated Raman scattering (SRS) microscopy has emerged as a powerful optical imaging technique for tissue diagnosis. In recent years, two-color SRS has been shown to be able to provide hematoxylin and eosin (H&E)-equivalent images that allow fast and reliable diagnosis of brain cancer. Such capability has enabled exciting intraoperative cancer diagnosis applications. Two-color SRS imaging of tissue can be done with either a picosecond or femtosecond laser source. Femtosecond lasers have the advantage of enabling flexible imaging modes, including fast hyperspectral imaging and real-time, two-color SRS imaging. A spectral-focusing approach with chirped laser pulses is typically used with femtosecond lasers to achieve high spectral resolution. Two-color SRS acquisition can be realized with orthogonal modulation and lock-in detection. The complexity of pulse chirping, modulation, and characterization is a bottleneck for the widespread adoption of this method. This article provides a detailed protocol to demonstrate the implementation and optimization of spectral-focusing SRS and real-time, two-color imaging of mouse brain tissue in the epi-mode. This protocol can be used for a broad range of SRS imaging applications that leverage the high speed and spectroscopic imaging capability of SRS.
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影响因子：
3.3
作者：
Fu, Dan;Holtom, Gary;Freudiger, Christian;Zhang, Xu;Xie, Xiaoliang Sunney
通讯作者：
Xie, Xiaoliang Sunney
影响因子：
3.4
作者：
Pence, Isaac J.;Kuzma, Benjamin A.;Evans, Conor L.
通讯作者：
Evans, Conor L.
DOI：
10.1073/pnas.1813044116
发表时间：
2019-04-02
影响因子：
11.1
作者：
Wei, Mian;Shi, Lingyan;Min, Wei
通讯作者：
Min, Wei
影响因子：
28.1
作者：
Zhang, Luyuan;Shi, Lingyan;Min, Wei
通讯作者：
Min, Wei
DOI：
10.1038/s41374-020-00514-0
发表时间：
2021-04
期刊：
Laboratory investigation; a journal of technical methods and pathology
影响因子：
--
作者：
Cui M;Zhang DY
通讯作者：
Zhang DY