雙光子光學顯微造影和高精密切片技術於肺腺癌三維結構影像重組之應用

Abstract

癌症，近年來高居全國十大死因之首，而肺癌嚴重性又是更甚於其中。本論文和實驗室同仁主要研究於利用新的三維結構組建平台、3D列印機、實驗室開發的高精準振動切片儀器、雙光子螢光顯微系統，把肺腺癌分化出來的兩種細胞株(Cell lines) CL1-0和CL1-5的腫瘤做三維結構重組和影像分析。過去研究都指向CL1-5較具有侵略性，所以我們希望藉由此論文可以改變傳統癌症上只考量到二維結構的病理結構分析。在近幾年來免疫系統和癌症的交互作用在醫學界中討論的如火如荼，因此癌症分析理論上應該多琢磨考慮三維的細節，此論文以三維影像重組定量分析找出較為適當的討論。建造並提出一整套系統可以將所有關於腦、肺、角膜、皮膚、肝…等等三維結構的影像重組，加上此論文發展出一套新的技術，可以直接把所有組織切片排成矩陣(tissue array)。染色完就可以直接在雙光子激發螢光顯微鏡下觀測，也可以節省大量的人力和時間成本。

Cancer is the most common cause of death in Taiwan in recent years and this includes lung cancer. We mainly investigated the use of a new 3-D structure construction platform, a 3-D printer, a high-precision vibrating microtome developed by the laboratory, and a two-photon microscopy system for 3-D structural reconstruction and image analysis of two differentiated lung adenocarcinoma cell lines, CL1-0 and CL1-5. Previous studies have consistently found that CL1-5 is highly invasive. We hope that the findings stated in this thesis can change the conventional 2-D structural pathological analysis of cancers. In recent years, there has been much discussion about the interactions between the immune system and cancer in the medical field; therefore, cancer analysis should theoretically consider 3-D details and we used 3-D image reconstruction and quantitative analysis to identify appropriate discussion points. The constructed and proposed system can carry out 3-D structural reconstruction of the brain, lungs, cornea, skin, liver, and other organs. In addition, a new technology was developed in this study; this technology can directly section all tissues into a tissue array, which can then be directly observed under a two-photon microscope after staining is complete. This can result in significant savings on personnel effort and time.