Shot-Noise-Limited Two-Color Stimulated Raman Scattering Microscopy with a Balanced Detection Scheme,The Journal of Physical Chemistry B

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Shot-Noise-Limited Two-Color Stimulated Raman Scattering Microscopy with a Balanced Detection Scheme
The Journal of Physical Chemistry B ( IF 2.8 ) Pub Date : 2020-03-24 , DOI: 10.1021/acs.jpcb.0c01065
Youngjin Choi _{1,

2} , Sohee Lim _{1,

2} , Joong Won Shim _{1,

2} , Bonghwan Chon ₁ , Jong Min Lim ₁ , Minhaeng Cho _{1,

2}

Affiliation

Stimulated Raman scattering (SRS) microscopy has been considered a useful technique for investigating chemical components by selectively targeting the vibration mode of chemical structures. Its practical application to the observation of molecular structures and dynamics in complicated biological environments requires broad spectral coverage with both high resolution and a high signal-to-noise ratio. Here, we demonstrate a two-color SRS microscopy employing a balanced detection scheme and a spectral focusing method. Two different SRS signals are generated with pump and Stokes laser pulse pairs in perpendicular polarization, where each of them acts as an intensity reference for the other, significantly reducing the background noise level close to the shot-noise limit even with a fiber-based femtosecond laser system. The high spectral resolution comparable to that of spontaneous Raman scattering spectroscopy is achieved with the spectral focusing method. The two-color SRS images are obtained for a mixture of polymer beads and for the distributions of lipids and proteins in U2OS cells.

中文翻译：

具有平衡检测方案的散粒噪声限制的两色受激拉曼散射显微镜

通过选择性地针对化学结构的振动模式，受激拉曼散射（SRS）显微镜被认为是研究化学成分的有用技术。它在观察复杂生物环境中的分子结构和动力学方面的实际应用需要具有高分辨率和高信噪比的宽光谱范围。在这里，我们演示了一种采用平衡检测方案和光谱聚焦方法的双色SRS显微镜。泵浦和斯托克斯激光脉冲对在垂直偏振状态下会产生两个不同的SRS信号，其中每个信号对彼此都作为强度参考，即使使用基于光纤的飞秒，也可以将背景噪声水平降低到接近散粒噪声极限激光系统。通过光谱聚焦方法可以实现与自发拉曼散射光谱法相当的高光谱分辨率。对于聚合物小珠的混合物以及U2OS细胞中脂质和蛋白质的分布，获得了两色SRS图像。

更新日期：2020-03-24

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