Fluorescence-Suppressed Raman Technique for Quantitative Analysis of Protein Solution Using a Micro-Raman Probe, the Shifted Excitation Method, and Partial Least Squares Regression Analysis

Authors: Oshima, Yusuke¹; Komachi, Yuichi²; Furihata, Chie³; Tashiro, Hideo⁴; Sato, Hidetoshi⁵

Source: Applied Spectroscopy, Volume 60, Issue 9, Pages 220A-236A and 951-1095 (September 2006) , pp. 964-970(7)

Publisher: Society for Applied Spectroscopy

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Abstract:

A practical Raman analyzing technique with suppression of the strong fluorescent background in order to obtain quantitative information is proposed in the present study. The technique is based on the shifted excitation method and partial least squares regression (PLSR) analysis. The Raman system consists of a single Raman spectrometer, a background-free electrically tunable Ti:Sapphire laser (BF-ETL), and a micro-Raman probe (MRP). The system allows one to obtain reliable shifted excitation Raman spectra with a simple operation. The PLSR analysis successfully provides quantitative information from the obtained spectra with the suppression of random noise including photon shot noise. The present study demonstrates that the technique is effective for extracting quantitative information concealed behind a fluorescent background that is more than 200 times stronger than the Raman signal.

Keywords: RAMAN SPECTROSCOPY; QUANTITATIVE ANALYSIS; TUNABLE LASER; RAMAN PROBE; FLUORESCENCE REJECTION; SHIFTED EXCITATION; CHEMOMETRICS

Document Type: Research article

DOI: 10.1366/000370206778397344

Affiliations: 1: Tashiro Probing Technology Laboratory, Riken (The Institute of Physical and Chemical Research), Hirosawa, Wako-shi, Saitama 351-0198, Japan; Department of Chemistry and Biological Science, College of Science and Engineering, Aoyama Gakuin Univers 2: Tashiro Probing Technology Laboratory, Riken (The Institute of Physical and Chemical Research), Hirosawa, Wako-shi, Saitama 351-0198, Japan; Machida Endoscope Co. Ltd., 6-13-8 Honkomagome, Bunkyo-ku, Tokyo 113-0021, Japan 3: Department of Chemistry and Biological Science, College of Science and Engineering, Aoyama Gakuin University, Fuchinobe, Sagamihara-shi, Kanagawa 229-8558, Japan 4: Tashiro Probing Technology Laboratory, Riken (The Institute of Physical and Chemical Research), Hirosawa, Wako-shi, Saitama 351-0198, Japan 5: Tashiro Probing Technology Laboratory, Riken (The Institute of Physical and Chemical Research), Hirosawa, Wako-shi, Saitama 351-0198, Japan; Optical Biopsy Development Research Unit, Riken (The Institute of Physical and Chemical Research), Hirosa

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