STUDY ON FT-IR MICROSCOPIC CHARACTERIZATION OF CELLS AND TISSUES IN PROTEIN SECONDARY STRUCTURE

• Main Authors: YUEH LEE, 李岳
• Other Authors: How Tseng
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/10899737416240526210

碩士 === 臺北醫學大學 === 醫學研究所 === 91 === At present the approach to diagnosis malignant tissues and cells all are by pathological section and use microscopy to observe their morphology. However, the traditional histopathological analysis still remaining in certain lacks and Fourier-transform infrared spectroscopy (FT-IR) employs a unique approach to optical diagnosis of tissue pathology based on the characteristic molecular vibrational spectra of the tissue, because of these advantages it is a powerful tool for diagnosis in medical field. Therefore it is important for our study to establish a pattern and from this pattern we can get novel and reasonable method for diagnosis of the cancer. By alternately compares with the changes of protein secondary structure in malignant and normal tissues and cells provides the deep rich insight the understanding to at present still not clear some cancer lines. The steps of this study (1) select the sample from malignant and normal tissues, (2) preparation and selection of the attachment (3) the FT-IR special absorption peak alternately compares (4) the changes of protein secondary structure in cells which administrate drugs or not. It provides an important reference for clinical cure or diagnosis, with systemic data base collection and analyses through the methods of frozen cut slide and FT-IR spectroscopy. In tissue, the spacemen embedding deposits as for -80OC for fresh-frozen, and prepare to 5mm thickness at -25OC for frozen -sectioning , then by FT-IR spectroscopy observes the changes of protein secondary structure in malignant and normal tissues. In cell, colon cancer cell line (HT29, Colo 205) and hepatoma (Hep G2, Hep 3B) which with attach character are selected, then attach on the prepared Ti disc, wafer and glass disc, by same way to get the information from the optical and IR spectra. Then the peaks were deconvoluted by the Peak Fit software. The purpose in this study is to compare the alternation of protein secondary structure between α-helix and β-type conformations in amide I region. In the tissue, the percentage of β-type conformation in tumoral tissue is higher than it''s in normal tissue, and there is a trend that the peak shift to the higher wavenumber. The result in the cell depends on the concentration of the EtOH which were treated to the cancer cell. There is a trend that peak shift to lower wavenumber in higher concentration than in the lower concentration of EtOH. In the future, we hope that FT-IR can be used in different diseases and compare with the normal one. Further more, find the regular role to use the information in the diagnosis or cure in the clinic.