受激拉曼散射是相干拉曼散射中的一种, 其产生的信号在三阶非线性效应下得到了显著地增强, 且没有非共振背景的干扰, 光谱与自发拉曼光谱几乎完全一致。 因此, 基于受激拉曼散射的显微成像技术具有无标记、 高特异性、 非侵入等优点, 已成功运用在生物细胞成像中并取得了许多重大的成就。 受激拉曼信号与激发光的波长相同, 易受到激发光背景噪声的干扰, 为解决该难题, 常采用光学调制与相敏检测相结合的方法对其进行检测。 检测过程中, 调制深度对受激拉曼信号强度和信噪比有重要影响。 针对此, 基于相关理论深入分析了调制深度对受激拉曼信号强度及信噪比的影响。 同时考虑到在生物光谱成像等应用中, 细胞光损伤阈值对两束激发光功率之和的限制, 分析了不同调制深度下, 获取最大信号强度及最佳信噪比的激发光功率配置方法。 通过搭建受激拉曼实验系统, 以二甲基亚砜为研究对象, 进行实验验证。 研究结果表明, 在光损伤阈值的限制条件下进行受激拉曼损耗检测时, 同一调制深度下, 当泵浦光与斯托克斯光光功率比为1: 1时信号强度达到最强, 比值为1∶2时信号的信噪比达到最佳。 在泵浦光与斯托克斯光光功率比相同的条件下, 受激拉曼信号强度和信噪比均随调制深度的降低而降低且近似呈线性相关。 实验得到的二甲基亚砜受激拉曼光谱图也验证了在实际样品的光谱检测中, 调制深度越高得到的光谱信号越强, 信噪比越佳, 整体的光谱质量也越好。 该研究结果是对受激拉曼显微技术在信号调制与检测方面的完善, 可为受激拉曼光谱检测和细胞成像实验做出参考性指导。

Stimulated Raman scattering is one of coherent Raman scattering. The signal generated by stimulated Raman scattering is significantly enhanced under the third-order nonlinear effect, and there is no interference from non-resonant background. Its spectrum is almost consistent with the spontaneous Raman spectrum. Therefore, the micro-imaging technology based on stimulated Raman scattering has the advantages of no labeling, high specificity and being non-invasive. It has been successfully used in biological cell imaging and has made many great achievements. Stimulated Raman signal has the same wavelength as excitation luminescence and is easily disturbed by excitation luminescence background noise. In order to solve this problem, the combination of optical modulation and phase-sensitive detection is often used to detect it. In the detection process, modulation depth influences the intensity and signal-to-noise ratio of the stimulated Raman signal. Because of this, this paper deeply analyzes the influence of modulation depth on stimulated Raman signal intensity and signal-to-noise ratio based on relevant theories. At the same time, considering the limitation of cell photodamage threshold on the sum of two excitation optical powers in applications such as bio-spectral imaging, the excitation optical power configuration method to obtain the maximum signal intensity and the best signal-to-noise ratio at different modulation depths is analyzed. By establishing a stimulated Raman experimental system, dimethyl sulfoxide is taken as the research object for experimental verification. The results show that when the stimulated Raman loss is detected under the limitation of photodamage threshold, at the same modulation depth, the signal intensity reaches the strongest when the optical power ratio of pump light to the one of stokes light is 1∶1,and the signal-to-noise ratio reaches the best when the ratio is 1∶2. When the optical power ratio of pump light to the one of stokes light is the same, the intensity and signal-to-noise ratio of stimulated Raman signal decrease with the decrease of modulation depth, and the correlation is approximately linear. The stimulated Raman spectrum of dimethyl sulfoxide obtained from the experiment also verified that the higher the modulation depth, the stronger the spectral signal and the better the signal-to-noise ratio and the better the spectral quality of the whole sample. The research results are the improvement of stimulated Raman microscopy in signal modulation and detection and can provide reference guidance for stimulated Raman spectroscopy detection and cell imaging experiments.