本研究提出一種可即時調控微電極結合磁場控制氧化鐵標記細胞排列於三維海藻酸鈣（Calcium Alginate）凝膠中。通過磁場的方向、強度及海藻酸鈣水凝膠的固化程度，仿天然組織器官在生物體外或體內培養細胞並排列，達到自組裝可調式組織結構。大部分人體器官都有特定的組織其建構在不同的細胞排列圖案以發揮各式功能，如人體的關節結締組織、心臟、肺臟、肝臟等。
本文先將大小1μm、濃度0.1~10mg/ml之MnZn軟磁性顆粒Ferromagnetic nano- or micro-particles （FMP）及BHK-21細胞共培養並混入海藻酸鈣水溶液中，藉強力磁鐵控制細胞形成仿人體器官組織之排列，另一方面磁粉也可增強整體組織建構性，接著使用晶片非晶矽氫（a-Si:H）照光後阻抗變小之特性，可形成真實電極的效果，取代傳統電化學所使用固定形狀之電極，並整合具有數位微型反射鏡元件（digital micromirror device, DMD）之投影機的光學架設，將電腦控制的影像經反射後透過物鏡聚焦成像在晶片上，成為可調變之微電極，電化學方式將溶液中碳酸鈣的鈣離子解離，解離出的鈣離子與海藻膠結合而凝膠化，達到固定膠體內部細胞之排列及外部海藻酸鈣水膠的位置和形狀。
合計上述的方法，此科技也適用於在生物體外基本生物學研究以及組織工程領域中製造複雜的仿生器官。

關鍵詞：鐵磁性顆粒、組織工程、非晶矽氫、數位微型反射鏡元件、可調控式微電極、海藻酸鈣水膠

This thesis presents iron oxide-labeled cells in three-dimensional calcium alginate gel using magnetic fields by the microelectrode of adjusting and controlling immediately. Through manipulating the strength, orientation of the magnetic field and using crosslinking gradients in calcium alginate gel，we assembly of cell-laden microgels to generate natural tissue of Tissue Engineering research constructs with tunable microarchitecture and complexity in vitro and even directly in situ.Most of human organs consist of multiple types of cells organized in a complex pattern to meet specific functional needs .Such as those of the articular cartilage heart, lungs, and liver.
We culture BHK-21 with MnZn ferromagnetic particles between 0.1 and 10mg/mL that was added in Alginate gels，by controlling magnets generate natural tissue，on the other hand，certain species of particles can permit or even enhance tissue formation . According to the characteristic of hydrogenated amorphous silicon (a-Si:H), which impedance diminished after light illuminating. Not only used light to create real electrodes, but also combined optics setup with DMD projector to be the microelectrode of adjusting and controling immediately. Electrode patterns can be changed quickly and efficiently, and this method replaces the traditional microelectrode especially. Therefore it can control the positions and forms of calcium alginate gel formation what we want.
Taken together, this technique can be adapted for studying basic biological processes in vitro and applicable for the fabrication of complex tissue architectures required in tissue engineering.
Key words : tissue engineering, a-Si:H, digital micromirror device, the microelectrode of adjusting and controling immediately, Calcium alginate

摘要 ……………………………………………………………………………Ⅰ
Abstract ………………………………………………………………………Ⅱ
圖目錄 …………………………………………………………………………Ⅵ
第一章 緒論 ……………………………………………………………………1
1.1前言 ……………………………………………………………………1
1.2文獻回顧 ………………………………………………………………5
1.2.1海藻膠水膠固化 ………………………………………………5
1.2.1.1外部固化法 …………………………………………….5
1.2.1.2內部固化法………………………………………………7
1.2.1.3光固化法 ………………………………………………8
1.2.1.4電化學固化法 …………………………………………10
1.2.2外部固化之3D細胞微陣列應用……………………………15
1.2.2 電化學固化之3D細胞微陣列應用………………………...17
1.2.3組織工程應用…………………………………………………21
1.3研究動機 ………………………………………………………….25
第二章 實驗原理………………………………………………………………26
2.1原理 …………………………………………………………………26
2.1.1電化學沉積海藻膠酸鈣原理 ………………………………26
2.1.2 非晶矽光電原理……………………………………………..26
2.1.3磁相關原理 …………………………………………………27
2.2實驗機制 …………………………………………………………29
第三章 實驗設計與架設 ……………………………………………………31
3.1實驗材料 ………………………………………………………….31
3.1.1晶片製作 ………………………………………………………31
3.1.2海藻膠 …………………………………………………………32
3.1.3細胞培養 ………………………………………………………33
3.1.4磁粒與細胞共培養 ……………………………………………33
3.1.5螢光染劑 ……………………………………………………34
3.2實驗流程 ………………………………………………………………36
3.3實驗架設 ………………………………………………………………39
第四章 實驗結果與討論 ……………………………………………………43
4.1實驗結果 ………………………………………………………………43
4.2實驗討論 ………………………………………………………………69
第五章 結論與未來展望 ……………………………………………………73
5.1結論 ……………………………………………………………………73
5.2未來展望 ………………………………………………………………74
參考文獻 ………………………………………………………………………76
附錄 …………………………………………………………………………79
7.1影像尺寸校正方法 ……………………………………………………80
誌謝……………………………………………………………………………82

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