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基于光纤探头的组织分析中,使用波长调制拉曼光谱进行荧光抑制。

Fluorescence suppression using wavelength modulated Raman spectroscopy in fiber-probe-based tissue analysis.

机构信息

University of St Andrews, SUPA, School of Physics & Astronomy, North Haugh, St Andrews, Fife, Scotland, KY16 9SS, United Kingdom.

出版信息

J Biomed Opt. 2012 Jul;17(7):077006. doi: 10.1117/1.JBO.17.7.077006.

Abstract

In the field of biomedical optics, Raman spectroscopy is a powerful tool for probing the chemical composition of biological samples. In particular, fiber Raman probes play a crucial role for in vivo and ex vivo tissue analysis. However, the high-fluorescence background typically contributed by the auto fluorescence from both a tissue sample and the fiber-probe interferes strongly with the relatively weak Raman signal. Here we demonstrate the implementation of wavelength-modulated Raman spectroscopy (WMRS) to suppress the fluorescence background while analyzing tissues using fiber Raman probes. We have observed a significant signal-to-noise ratio enhancement in the Raman bands of bone tissue, which have a relatively high fluorescence background. Implementation of WMRS in fiber-probe-based bone tissue study yielded usable Raman spectra in a relatively short acquisition time (∼30  s), notably without any special sample preparation stage. Finally, we have validated its capability to suppress fluorescence on other tissue samples such as adipose tissue derived from four different species.

摘要

在生物医学光学领域,拉曼光谱是探测生物样本化学成分的有力工具。特别是,光纤拉曼探头在体内和体外组织分析中起着至关重要的作用。然而,组织样本和光纤探头的自发荧光通常会产生很强的高荧光背景,这强烈干扰了相对较弱的拉曼信号。在这里,我们展示了波长调制拉曼光谱(WMRS)的实现,该技术可以在使用光纤拉曼探头分析组织时抑制荧光背景。我们已经观察到骨组织中具有相对较高荧光背景的拉曼带的信噪比显著增强。在基于光纤探头的骨组织研究中实现 WMRS,在相对较短的采集时间(约 30 秒)内获得了可用的拉曼光谱,特别是没有任何特殊的样品制备阶段。最后,我们验证了它在其他组织样本(如来自四个不同物种的脂肪组织)上抑制荧光的能力。

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