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.
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.
Oshima
Yusuke
Y
Tashiro Probing Technology Laboratory, Riken (The Institute of Physical and Chemical Research), Hirosawa, Wako-shi, Saitama 351-0198, Japan.
Komachi
Yuichi
Y
Furihata
Chie
C
Tashiro
Hideo
H
Sato
Hidetoshi
H
D000818:Animals; D002417:Cattle; D002645:Chickens; D005453:Fluorescence; D016018:Least-Squares Analysis; D009113:Muramidase / Q000032:analysis*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000032:analysis*; D012996:Solutions; D013059:Spectrum Analysis, Raman / Q000295:instrumentation / Q000379:methods
D012996:Solutions; D012710:Serum Albumin, Bovine; D009113:Muramidase