Application of Raman Micro-spectroscopic and Fourier transformed Infrared Micro-spectroscopic Techniques for Clinical Test.

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(1) FT-IR microscopic imaging of frosen-sectioning experimental tumor tissue :Our purpose is to develop infrared (IR) microspectroscopy as a new optical diagnostic tool to support conventional lightscopic techniques in investigating the viability of carcinoma tissues and to develop its use in the evaluation of the early effects of anticancer therapy by monitoring the IR spectra in the necrotic area.We ammased for 4 weeks after the isotransplantation of mouse squamoouscell carcinoma into the thigh of mice. The borders of the necrotic area of frozen tissue specimens were investigated by Fourier-transform IR microspectroscopy and conventional histological staining.In the results, a significantly higher accumulation of cholesterol was observed in the necrotic tissue of a carcinoma. The mechanism of this phenomenon is hitherto unrecognized. We proposed that the accumulated cholesterol may lie extracellularly as a result of the ruptured plasma and internal membranes after the swelling of the … More necrotic cells brought on by hypoxia. The analysis of the secondary structure of protein revealed that the amount of β-sheet increased significantly in striking contrast to the decreasing amounts of α-helix in a necrotic area of a carcinoma. It is plausible that this structural conversion of protein was because of the liquid-autooxidation products, such as cholesterol oxide but not cholesterol itself, which processes cell toxicity and could be generated in a necrotic area.(2) Raman microscopic imaging of hard tissues (Calcified Cartilage) :The aim of the present study was to investigate the quantity and quality of hydroxyapatite (HA) in the zone of calcified cartilage of articular cartilage and underlying subchondral bone by Fourier transform infrared microscopy (FTIRM).The mineral:matrix ratio was determined by integrating areas of υ1, υ3 bands at 900-1200cm^<-1> to Amide I band at 1585-1725cm^<-1>. The CO_3:PO_4 ratio was determined by integrating areas of υ2 carbonate band at 850-900cm^<-1> to υ1. υ3 bands. In the control, the mean mineral:matrix ratio of the medial side was significantly lower than the lateral side at the zone of the calcified cartilage and subchondral bone. The mean CO_3:PO_4 ratio was significantly higher in the medial side than the lateral one at the zone of the calcified cartilage and subchondral bone. In contrast, the results obtained from the model showed no significant differences between the two sides. Our results showed diminished deposition and increased maturation of HA in control. These findings may correlate with the pathological process of knee OA, and indicate that FTIRM is a useful tool for determining the variability of HA in OA. Less