Wang Zhiqiang, Ju Siwen, Zhou Xiaofei, Ni Feng, Qiu Yanhua, Zhang Ruiting, Ma Lin, Lin Ke
School of Physics, Xidian University, Xi'an, 710071, P. R. China.
The Affiliated Hospital of Xidian University, Xi'an, 710071, P. R. China.
Anal Bioanal Chem. 2024 Nov;416(28):6259-6267. doi: 10.1007/s00216-024-05538-9. Epub 2024 Sep 18.
Raman spectroscopy is an important technique for analyzing the chemical composition of samples in many fields. A severe challenge often encountered in Raman measurements is the presence of a concurrent fluorescence background, especially in biological samples. In order to obtain accurate Raman spectra, the fluorescence background must be subtracted from the original Raman spectra. We proposed a shifted ratio spectrum method to subtract the strong fluorescence background from the original Raman spectrum. First, the original Raman spectrum is divided into multiple regions according to the spectral shape of the shifted ratio spectra, and then, Gaussian fitting is performed in each region. The fitting results are stitched together in order to obtain the complete fluorescence background. Finally, this fluorescence background is subtracted from the original spectrum to obtain a pure Raman spectrum. This method can accurately subtract the fluorescence background of Rhodamine 6G (R6G)/ethanol solution and serum. This highlights the great potential of this method for applications in both biological and non-biological samples.
拉曼光谱法是分析许多领域中样品化学成分的一项重要技术。拉曼测量中经常遇到的一个严峻挑战是同时存在荧光背景，尤其是在生物样品中。为了获得准确的拉曼光谱，必须从原始拉曼光谱中减去荧光背景。我们提出了一种移位比光谱法，用于从原始拉曼光谱中减去强荧光背景。首先，根据移位比光谱的光谱形状将原始拉曼光谱划分为多个区域，然后在每个区域进行高斯拟合。将拟合结果拼接在一起以获得完整的荧光背景。最后，从原始光谱中减去该荧光背景以获得纯拉曼光谱。该方法可以准确地减去罗丹明6G（R6G）/乙醇溶液和血清的荧光背景。这突出了该方法在生物和非生物样品应用中的巨大潜力。
