Development of an Ultraviolet Raman Microspectrometer Applicable to Structural Analyzes of Cells

适用于细胞结构分析的紫外拉曼显微光谱仪的研制

基本信息

批准号：
07554025
负责人：
TAKEUCHI Hideo
金额：
$ 10.18万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07554025/
关键词：
Microscope Protein Nucleic acid Cell structure UV resonance Raman spectroscopy

项目摘要

Raman microspectroscopy is a useful technique to investigate the structures of micrometer-sized particles in a non-destructive manner. On the other hand, ultraviolet resonance Raman spectroscopy provides key information on the structures of proteins and nucleic acids. In this study, we have developed an ultraviolet Raman microspectrometer to exploit the utilities of both spectroscopic methods. The microspectrometer consists of a continous-wave ultraviolet laser, an ultraviolet microscope, and a Raman spectrometer. The ultraviolet laser beam is obtained by frequency-doubling visible laser light from an Ar^+ laser. The ultraviolet microscope is specially designed for high throughput in the ultraviolet region and high spatial resolution of microscopic images. Samples under a microscope objective are irradiated with ultraviolet laser light and the Raman scattered light was collected with the same objective and then guided to the Raman spectrometer, which has already been developed in our laboratory. We have succeeded to record some Raman spectra by using newly developed ultraviolet Raman microspectrometer.However, the sensitivity of the microspectrometer is not enough to investigate the structures of microparticles in living cells. We are planning to use an ultra-high-sensitivity CCD detector to detect even extremely weak Raman light and to apply the ultraviolet Raman microspectrometer to structural analyzes of living cells.

拉曼显微光谱是一种以非破坏性方式研究微米尺寸颗粒结构的有用技术。另一方面，紫外共振拉曼光谱提供了蛋白质和核酸结构的关键信息。在这项研究中，我们开发了一种紫外拉曼显微光谱仪来利用这两种光谱方法的效用。显微光谱仪由连续波紫外激光器、紫外显微镜和拉曼光谱仪组成。紫外激光束是通过对 Ar^+ 激光器发出的可见激光进行倍频而获得的。紫外显微镜专为紫外区域的高通量和显微图像的高空间分辨率而设计。用紫外激光照射显微镜物镜下的样品，用同一物镜收集拉曼散射光，然后引导至我们实验室已经开发的拉曼光谱仪。我们利用新研制的紫外拉曼显微光谱仪成功记录了一些拉曼光谱。然而，显微光谱仪的灵敏度不足以研究活细胞内微粒的结构。我们计划使用超高灵敏度CCD探测器来检测极微弱的拉曼光，并将紫外拉曼显微光谱仪应用于活细胞的结构分析。