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Development and applications of highly-sensitive near-infrared multichannel Raman Spectroscopic system

高灵敏近红外多通道拉曼光谱系统的研制与应用

基本信息

批准号：
12354007
负责人：
HAMAGUCHI Hiro-o
金额：
$ 28.35万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

A highly sensitive near-infrared multichannel Raman spectroscopic system has been constructed. We use the 1064 nm line of a cw Q-switched Nd : YAG laser for excitation and a new InP/InGaAsP intensified CCD detector for detection. The system has a detectivity at least one order of magnitude higher than the conventional FT-Raman spectrometer and is capable of recording high S/N Raman spectra without the interference from fluorescence. 1) We are now able to measure the Raman spectra of human lung tissues, by virtue of fluorescence-free near-infrared excitation. We have found that the Raman spectra of cancered lung tissues are quite different from those of normal lung tissues. This result indicates that Raman spectroscopy distinguishes cancered and normal tissues very clearly and that it has a huge potential for in vivo and non-invasive clinical diagnosis of cancers. An optical-fiber irradiation/collection optics has been developed in order to facilitate in vivo Raman measurements. 2) Time- and space-resolved Raman spectra of a living yeast cell have been observed for the first time, using a confocal Raman microspectroscopic system with 632.8 nm excitation. Time- and space-dependent Raman bands of proteins, lipids, and sugars are observed, reflecting the evolution with the cell cycle of the molecular compositions in organelles like nucleus, mitochondrion and septum. 3) The near-infrared Raman system has been applied for the measurements of various protein and nucleic acid samples. Despite the fact that those samples fluoresce strongly with visible excitation, no interference from fluorescence has been observed with 1064 nm excitation. Structural change of DNA with drug intercalation has successfully been probed. In conclusion, the present research has established the methodology of near-infrared Raman spectroscopy, which is particularly useful in biomedical applications.

构建了高灵敏度近红外多通道拉曼光谱系统。我们使用 1064 nm 线的连续 Q 开关 Nd: YAG 激光器进行激发，并使用新型 InP/InGaAsP 增强型 CCD 探测器进行检测。该系统的探测灵敏度比传统的傅里叶变换拉曼光谱仪至少高一个数量级，并且能够记录高信噪比的拉曼光谱，而不受荧光的干扰。 1）我们现在能够通过无荧光的近红外激发来测量人体肺组织的拉曼光谱。我们发现癌变肺组织的拉曼光谱与正常肺组织的拉曼光谱有很大不同。这一结果表明拉曼光谱可以非常清楚地区分癌组织和正常组织，在癌症的体内和非侵入性临床诊断方面具有巨大的潜力。为了促进体内拉曼测量，已经开发了光纤照射/收集光学器件。 2) 使用 632.8 nm 激发的共焦拉曼显微光谱系统，首次观察到活酵母细胞的时间和空间分辨拉曼光谱。观察到蛋白质、脂质和糖的时间和空间依赖性拉曼带，反映了细胞核、线粒体和隔膜等细胞器中分子组成随细胞周期的演变。 3）近红外拉曼系统已应用于多种蛋白质、核酸样品的测量。尽管这些样品在可见光激发下发出强烈的荧光，但在 1064 nm 激发下没有观察到荧光干扰。已成功探测到药物嵌入引起的 DNA 结构变化。总之，本研究建立了近红外拉曼光谱的方法，该方法在生物医学应用中特别有用。