臺灣博碩士論文加值系統

蟹殼上之殼糖晶型結構、為生物型液晶之最佳研究材料。因具有規則的排列角度，並可於電子顯微鏡下觀察。本研究環繞殼糖為主軸、此種具有高分子特質之天然生物醫學材料；藉由蟹殼、烏賊軟骨、靈芝這三種樣品為實驗標的，期望在穿透式電子顯微鏡（TEM）底下搜尋膽酯醇型液晶相（cholesteric liquid crystal）的存在位置與構型。將蟹殼（crab）以2N NaOH去蛋白、2N HCl 脫鈣、冷凍乾燥脫水、EPON812平板包埋等程序處理後，於TEM 超薄切片（80nm）下證實 chitin cholesteric liquid crystal texture 的存在（Arced pattern of fingerprint）。後期將純化後之殼糖以3N HCl；以及不同濃度之NaOH， 104°C煮沸回流約1~6小時，離心稀釋及透析後將酸移除，並以超音波震盪後，使得降解過後之殼糖分子轉變為凝膠狀懸浮液，濃縮至一定濃度後產生分層現象，其分子具有類似節肢動物中殼糖微纖維的排列特質。簡言之，本研究的主要目的為搜尋膽酯醇型液晶並探討生体高分子液晶自我重組的特點。

The chitin crystalline architecture on crabs were by far the most suitable biomaterials for searching in liquid crystals. Owing to its regularly orientations and could be observed under transmission electron microscopy. The work here aims to chitin which is the biopolymers in nature, together with crabs, squid bone, and ganoderma residue as our targets for experiments. We try to find out the position and arrangement of chloesteric liquid crystal under transmission electron microscopy. First, the crab was treated by 2N NaOH to hydrolyze the protein, dissolved the mineral phase in 2N HCl, dehydrated, embedded and cured in Epon812, the arced fingerprint pattern in chitin thus could be viewed via microtone ultracut (80nm). Microfibrillar fragments of purified crab chitin were prepared by hydrolysis in 3 M HCl at its boilong point (104°C), together with NaOH at various concentration. After discarding the acid by centrifugal washing and dialysis, an ultrasound treatment converts the residual product to a colloidal suspension stabilized by NH3+ charges. When dehydrated to a critical concentration, spontaneous formation of a two-phase equilibrium system occurs. The upper phase (lower concentration) is isotropic and the lower phase is anisotropic. The latter displays chiral nematic order and dries to a solid film which mimics the helicoid organization characteristic of the chitin microfibrils in the cuticle of arthopods.

目 錄
致謝………………………………………………………………………………….Ⅰ
中文摘要…………………………………………………………………………….Ⅱ
英文摘要…………………………………………………………………………….Ⅲ
目錄………………………………………………………………………………….Ⅳ
圖表目次…………………………………………………………………………….Ⅵ
第一章緒論……………………………………………………………………….1
1.1 前言………………………………………………………………………….1
1.2 研究目的…………………………………………………………………….6
第二章文獻回顧………………………………………………………………….7
2..1 生物液晶之起源……………………………………………………………7
2.1.1 生物組織結構的疑問………………………………………………..7
2.1.2 細胞組織的流動性及次序…………………………………………..8
2.2 液晶性質和相似物………………………………………………………….9
2.2.1 Intermediate State of Matter…………………………………………..9
2.2.2 液晶的分類…………………………………………………………10
2.2.3 液晶之生物同源性…………………………………………………11
2.3 膽固醇幾何構型之性質與推論…………………………………………..12
2.3.1 Twisted Plywood Model…………………………………………….12
2.3.2 Periodic Extinctions in Polarized Light……………………………..13
2.3.3 Arced Patterns in Transmission Electron Microscopy………………14
2.3.4 Deviations, Defects, and Artifacts…………………………………..15
第三章研究材料與方法………………………………………………………...17
3.1 材料與試劑………………………………………………………………...17
3.2 儀器設備…………………………………………………………………...18
3.3 研究方法及進行步驟……………………………………………………...20
3.3.1 穿透式電子顯微鏡…………………………………………………20
3.3.2 掃描式電子顯微鏡…………………………………………………22
3.3.3 偏光顯微鏡…………………………………………………………23
3.3.4 廣角X-ray繞射分析……………………………………………….24
3.3.5 粘度測定……………………………………………………………25
3.3.6 顯微鏡式傅利葉轉換紅外光譜分析………………………………25
3.3.7 示差掃描式熱分析儀………………………………………………27
3.3.8 元素分析……………………………………………………………27
3.3.9 CP MAS 13C NMR ………………………………………………….28
3.3.10微電場對自我重組之影響………………………………………...28
第四章結果與討論……………………………………………………………...29
4.1 穿透式電子顯微鏡………………………………………………………...29
4.2 掃描式電子顯微鏡………………………………………………………...31
4.3 偏光顯微鏡………………………………………………………………...32
4.3.1 光學顯微鏡下之超薄切片…………………………………………33
4.3.2 colloids形成及回收率………………………………………………33
4.3.3 Phase separation of biphasic system………………………………...33
4.3.4 The role of the pH in the formation of cholesteric liquid crystal spherulites from a-chitin…………………………………………..35
4.3.5 Initiation of the spherulites and the effect of temperature on spherulite formation…………………………………………………………..37
4.3.6 弧狀指紋構形………………………………………………………40
4.4 X-ray繞射分析……………………………………………………………..44
4.5 還原黏度測試……………………………………………………………...45
4.6 IR圖譜分析………………………………………………………………...46
4.7 DSC熱性質分析……………………………………………………………47
4.8 EA元素分析………………………………………………………………..48
4.9 CP MAS 13C NMR………………………………………………………….49
4.10微電場對自我重組之影響………………………………………………..49
第五章結論……………………………………………………………………...51
參考文獻……………………………………………………………………………..53
註……………………………………………………………………………………..92
附錄…………………………………………………………………………………..94

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