臺灣博碩士論文加值系統

多光子顯微術使用近紅光作為激發光源、具有描掃深度深、可激發所有可見光波段的螢光及二倍頻訊號的優點使樣品不需染色; 自動光學切片使樣品不需切片並可沿深度掃瞄得到三維影像,是一種低侵入性生物三維造影技術。是非常有力的工具。我們可以把應用於生物可分解性的材料及正常與癌症組織的初步診斷。

第一部分 :生物可降解性的材料
組織工程為組織修復及器官複製帶來新希望，其所用的材料為生物可降解性。在此篇論文中，我們先嘗試使用多光子顯微術來觀測這些材料的非線性光學特性，作用未來鷹架與細胞間的交互作用的對照組。結果發現以上所掃瞄的生物可分解的薄膜的光譜多偏向藍光，膠原蛋白鷹架在水中會降解而使得二倍頻訊號降低等。

第二部分: 正常與癌症組織的初步診斷
鼻咽部位於鼻部與咽部90 度交會的隱密地帶。鼻咽癌乃東南華人特有之癌症。傳統診斷方式是組織病理切片, 製作需費時2 小時~1 天及需要醫生的長期經驗來判斷。於是我們利用多光子顯微術的優點，希望能藉此快迅區分出鼻咽組織中的正常細胞與癌細胞。並可做到傳統病理染切片染色難做到的定量分析。
實驗結果對於NPC 沒有區分出正常細胞及癌細胞，原因在於鼻咽部細胞種
類眾多，特別的是NPC 癌細胞會與正常細胞夾雜。
另一種對國人健康造成威脅是腎臟癌(Renal cell carcinoma)，我們可以使用二倍頻對自體螢光的指數及強度分佈圖等來區分正常腎組織及癌症腎組織。

Multiphoton microscopy, which uses near infrared light as excitation light source,has deeper penetration depth, and automatic optical sectioning ability. It is a minimally
invasive bioimaging technique and can capable visible autofluorescences excitation. Thespecimens used for multiphoton microscopy need not be sliced and dyed, and the imagesobtained clearly displays cell morphology, collagen, and elastins. In this thesis, we apply
multiphoton and second harmonic generation microscopy to biodegradable materialimaging and to cancer diagnosis.
Part I: Biodegradable scaffolds and films
Tissue engineering brings new hope for tissue recovery and organ replication. One important aspect of tissue engineering is utilizing suitable biodegradable material as a scaffold. We observed in these biomaterial’s nonlinear optical properties. We found that most biodegradable materials emit blue light. Therefore, if the cell’s autofluorescence is weak, green or red fluorescence protein should be chosen to transfect
the cells.
Part II：Preliminary diagnosis of Normal and cancerous tissue
Nasopharynx is the concealed region where the pharynx and the nose intersect each other perpendicularly. Nasopharyngeal carcinoma(NPC) is a type of cancer peculiar to southeaster Chinese. The traditional method of diagnosing nasopharyngeal carcinoma is with histology, which can be time consuming to prepare.
We attempted to utilize multiphoton and second harmonic generation microscopy to distinguish normal nasopharyngeal cell from nasopharyngeal carcinomatous cell. We
find that multiphoton microscopy cannot effectively make the distinction. The reason is that naspharyngeal cells are numerous, and the cancer cells of NPC are mixed with the normal cells.
Kidney cancer (Renal cell carcinoma) is another cancer that threatens the health of Chinese people. In this case, we were able to use an index of second-harmonic generation
to autofluorescence and an intensity distribution diagram to distinguish the normal kidney tissue from cancerous kidney tissue.

目錄…………………………………………………………I
致謝…………………………………………………………Ⅲ
英文摘要……………………………………………………Ⅳ
中文摘要……………………………………………………Ⅴ
圖表目錄……………………………………………………Ⅵ
1 緒論1
1.1 組織工程與生物可分解性材料簡介1
1.2 癌症研究的重要性2
2 傳統醫學檢驗方法7
2.1 非光學診斷法7
2.2 光學顯微術與組織切片8
3 非線性光學顯微術13
3.1 單光子與雙光子吸收15
3.2 倍頻產生的機制19
4 多光子顯微術儀器23
4.1 顯微鏡之前的光處理23
4.2 顯微系統與光子資訊處理25
5 生物可分解性材料的非線性光訊號
5.1 材料選擇27
5.2 材料的非線性光學反應28
6 區分鼻咽癌組織與正常組織
6.1 鼻咽及鼻咽癌簡介39
6.2 實驗方式與結果討論40
7 腎細胞上皮癌組織與正常腎上皮組織之
非線性顯微影像
7.1 腎及腎細胞上皮癌簡介47
7.2 光訊號飽和測試48
7.3 量化分析及圖表比較50
7.4 傳統H.E.染片結果62
7.5 結果解釋62
8 結論64
附錄:上皮組織65
參考文獻66

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