- 學位論文
全域式光學同調斷層掃描術結合拉曼光譜儀用於生物樣本的特性分析
Full-field Optical Coherence Tomography Combined with Raman Spectroscopy for Biological Sample Characterization
摘要
檢測完整的生物組織特性需要樣本的形貌資訊與生物化學資訊，光學同調斷層掃描術(Optical coherence tomography; OCT)擅於呈現樣本組織的立體微結構，但卻缺少生物分子的資訊；拉曼光譜儀(Raman spectroscopy)可以提供組織的生化成分資訊，但卻無法呈現組織的立體微結構資訊。為了能獲得完整的生物樣本資訊，在本論文中，我們將研發能獲得生物樣本立體形貌與生化成分資訊的拉曼光譜儀結合全域式光學同調斷層掃描術系統，並展示此系統應用於in vitro細胞株與ex vivo皮膚組織之量測結果。 在in vitro黑色素癌細胞的量測上，首先使用OCT系統建構出黑色素癌細胞的立體影像與其在空間中的位置，接著再利用此OCT影像引導拉曼光譜儀在細胞的位置上進行拉曼光譜的採集。實驗結果顯示，OCT的立體影像可以忠實地呈現細胞的立體形貌、實際大小(約10–15 μm)以及在空間中的具體位置，此外，拉曼頻譜也確實記錄到黑色素癌細胞的拉曼特徵訊號(1326 cm-1、1469 cm-1、1660 cm-1、2939 cm-1)，其拉曼峰值位置與文獻相符。 在ex vivo皮膚組織的量測上，分別對黑色素細胞癌組織蠟塊切片與正常皮膚組織蠟塊切片進行量測與區分，實驗結果顯示，雖然無法單純從兩者的OCT立體影像進行型態鑑別，但是兩者的拉曼頻譜有存在著差異，其差異在於有無位於1326 cm-1的拉曼訊號，因此能利用此拉曼訊號進行黑色素細胞癌組織與正常皮膚組織的辨別。 本論文展現拉曼光譜儀結合全域式光學同調斷層掃描術系統應用於量測生物樣本的潛力，不僅能獲得生物樣本的立體形貌，還能得知樣本所含有的生物分子資訊，讓生物樣本的分析得以更加地完整，以利於醫生對於病灶的診斷。
並列摘要
Complete characterization of biological tissue requires probing the morphological and biochemical information. Optical coherence tomography (OCT) excels at presenting three-dimensional (3-D) images of tissue microstructure but lacks biomolecular information, while Raman spectroscopy is capable of providing tissue biochemical composition with superb specificity, but cannot gather 3-D microstructure. In order to obtain the complete biological sample information, in this work, we develop an integrated full-field OCT (FF-OCT) and Raman spectroscopy system capable of acquiring 3-D morphology and biochemical composition from biological samples, and demonstrate the capabilities of this system with in vitro cell line, as well as ex vivo human skin tissue. In the in vitro melanoma cell line measurement, first, our FF-OCT system is used to construct the 3-D image of melanoma cells and acquire their position in space. Then, we utilize this 3-D image to guide the acquisition of Raman spectrum from a localized melanoma cells. The experimental results show that 3-D OCT images can truly present the 3-D morphology, size (10–15 μm) and specific position in space of cells. Furthermore, the characteristic Raman peaks of melanoma cells (1326 cm-1、1469 cm-1、1660 cm-1、2939 cm-1) are indeed detected by Raman spectroscopy, which is consistent with the results found in literature. In the ex vivo experiment, we measure the melanoma biopsy and normal skin biopsy, and try to discriminate between them by the integrated FF-OCT and Raman spectroscopy system. The experimental results show that it cannot simply distinguish between melanoma biopsy and normal skin biopsy by the OCT image, but there is also a difference in the Raman spectrum at 1326 cm-1, enabling us to differentiate melanoma skin tissue and normal skin tissue by this Raman signal. This work shows the potential of applying integrated FF-OCT and Raman spectroscopy system to biological tissue measurements. By using this system, not only the 3-D tissue microstructure can be obtained, but also the biochemical composition, resulting in a more complete analysis of biological samples, which provides better assistance for the diagnose of diseases.