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  • 學位論文

全域光學同調顯微術於上皮細胞與組織之逆散射量測與分析

Measuring and Analyzing Backscattering Intensity of Epithelial Cells and Tissues by Full-Field Optical Coherence Microscopy

指導教授 : 宋孔彬

摘要


人類癌症的產生常源自上皮層,目前已有許多文獻探討並指出上皮細胞的癌病變過程,會伴隨內部結構與折射率之變化,即直接影響逆散射光的變化;組織方面,上皮層與基質層的界線會漸模糊難辨,且有許多研究團隊量測並萃取組織的散射係數(scattering coefficient: μs)來做為診斷癌變之參考依據。因此,建立一套能夠量取逆散射光的技術,於生醫工程的發展與應用上實為一備具潛能的利器。此研究主要希望能建立一套光學實驗系統及方法,來對正常及癌變的口腔上皮細胞與口腔、食道組織作逆散射光強之量測與收集,進而將之分析並尋找出區別與診斷之依據。 目前採用之光學系統為「全域光學同調顯微術(Full-field Optical Coherence Microscopy, FF-OCM)」,主要根據低同調干涉理論來擷取樣本三維切面之逆散射影像。已完成系統響應之量測與改善、強反射面與小球仿體之逆散射影像還原、不同濃度小球仿體其隨深度衰減之比例分析,及多種溶液之平均折射率量測…等。現在正著手進行口腔正常細胞(SG cells)與癌細胞(CA9-22 cells)的逆散射光強分析,以及利用醋酸溶液輔助觀察分析之實驗。組織方面,冷凍切片已能利用系統觀察其內部型態與構造,唯背景干擾及切片本身散射光強之擷取,將尋找更佳之改善方法;塊狀組織目前可還原深度剖面之散射分布情形,然上皮層與基質層之確認尚在努力中,並初步對逆散射光強隨深度分布與衰減情形進行分析與探討。 結果已能定量確認CA9-22 cells添加0.4%醋酸溶液後,其逆散射光強會遠大於SG cells,且散射來源大部分來自細胞核,可作為診斷之參考資訊;組織方面衰減程度的分析,因量測樣本數目尚少僅處測試階段,期未來能更進一步針對組織之診斷有所發現與貢獻。

並列摘要


Human cancer is often generated from epithelium. Recently, many researches indicate that carcinoma in epithelial cells are accompanied with changes of structure and refractive index. It may also affect the backscattering intensity of cells directly. And when it comes to cancerous tissue, the boundary between epithelium and basal layer is gradually blurred. Many research teams has aimed for measuring and extracting the scattering coefficient(μs) in order to find out and diagnose the existence of carcinoma. Therefore, establishing the technique able to derive the backscattering light is a promising tool in the development and application of biomedical engineering. The goal of the thesis is to build the optical system and some methods that could collect the backscattering light from normal or cancerous cells and tissues. We want to find out how to distinguish and diagnose healthy cells/tissues and cancerous ones. The optical setup in this thesis is “Full-Field Optical Coherence Microscopy, FF-OCM.” It is based on the low coherence interferometry for extracting the 3D backscattering images of the sample. Presently, the measurement and improvement of the system response, the backscattering images reconstruction of the strong reflective planes and polystyrene bead phantom, the analysis of backscattering intensity decayed with the depth of the polystyrene phantoms in different ratios, and the refractive index measurements of many solutions have been finished. And analyzing the backscattering intensity of normal(SG cells) and cancerous(CA9-22 cells) oral epithelial cells before/after addition of acetic acid is still proceeding. On the tissue experiment side, frozen tissue sections are being looked for more perfect way for reconstruction of the backscattering images; The longitudinal backscattering distribution of bulk tissues have been rebuilt. However, the changes of the backscattering intensity along the depth of tissue are preliminary being analyzed. The results show that the backscattering intensity of CA9-22 cells would be much stronger than SG cells after addition of 0.4% acetic acid. Most of the backscattering light is from the nuclei of cells. It could be used as the diagnostic reference. The analysis of tissues is still in the testing process because of too small amount of the data. In the future, it is hoped that we can find out the way to diagnose normal and cancerous tissues.

參考文獻


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