本研究分為兩大主題，其一為建立灰階影像分析系統用於檢測細胞四氧化三鐵磁流體之磁標定量，並以其為檢測平台與細胞磁泳及比色分析法之定量結果比較；另一主題以細胞磁泳技術分析細胞膜特性改變後，對於細胞於液體中移動時之黏滯力的影響。第一個研究主題以人類肝癌細胞HepG2為分析模型，改變磁流體與細胞共培養之時間與濃度等參數後，進行細胞攝取量之定量分析。研究結果顯示，細胞磁泳以及灰階影像分析法皆適用於量測單顆細胞之磁標定量；比色分析法則可快速檢測一群細胞之平均鐵含量。而進行探討細胞膜表面差異之研究主題中，以人類肝癌細胞SKHep-1、老鼠纖維母細胞NIH-3T3及老鼠巨噬細胞RAW 264.7三株細胞進行實驗，分別以過氧化氫造成細胞膜損傷及以不同蛋白質包覆細胞外部後進行磁泳試驗，結果顯示兩種處理方式皆會大幅影響磁泳速度。經過比色法及灰階影像分析結果確定，細胞之磁標定量在經過處理後無明顯差異。而細胞粒徑分析結果亦顯示，表面處理不影響細胞整體球狀形態及尺寸。上述試驗可確認磁泳速度之變化主要來自於細胞膜特性改變，進而影響細胞泳動過程中之細胞黏滯力。

The aims of the study are to (1) build up an image analysis system for quantifying the intracellular iron oxide nanoparticles in labeled cells by measuring the grey-value and (2) to detect the alteration of cell membrane surface via cell magnetophoresis. For the first topic, the human hepatocellular carcinoma cell HepG2 was used as a cell model and both concentration- and time- dependent iron uptake were investigated. The results showed the cell magnetophoresis and the grey-value analysis were able to quantify the magnetic labelling of a single cell while the colorimetric assay was suitable for quick detection of large sample size. In the other topic, hepatocellular carcinoma cell SKHep-1, mouse fibroblast NIH-3T3, and mouse macrophage RAW 264.7 were used. Cells were treated by either hydrogen peroxide to induce oxidative damage or wrapped by protein. Significant differences were observed in cell velocities from two groups during magnetophoresis. The results from colorimetric assay and optical microscope indicated that intracellular MNP remained unchanged and cells maintained their overall morphology after treatment, which indicated that the differences between cellular velocities were caused by the alteration in cell membrane properties that affected the viscosity force during magnetophoresis.

摘要 I
Abstract II
目次 III
圖目次 VI
表目次 IX
第一章 緒論 1
1-1 研究背景與動機 1
1-2 研究目的 2
第二章 理論基礎與文獻回顧 3
2-1 磁性理論 3
2-1-1 磁性奈米顆粒 4
2-1-2 磁流體 5
2-2 細胞磁標定及生醫應用 6
2-2-1 磁性分離 7
2-2-2 藥物傳輸 8
2-2-3 磁振造影增強 9
2-2-4 磁熱治療 9
2-2-5 免疫檢測 11
2-3 細胞磁定量 12
2-3-1 磁泳定量 13
2-3-2 比色分析法 15
2-3-3 灰階影像分析 15
2-4 細胞表面構造與性質 17
第三章 實驗材料與方法 19
3-1 實驗架構 19
3-2 實驗材料 20
3-3 實驗設備 22
3-4 實驗方法 22
3-4-1 細胞培養 22
3-4-2 細胞磁標定 24
3-4-3 細胞表面性質調整 25
3-4-4 細胞磁泳分析 26
3-4-5 比色分析定量 26
3-4-6 灰階影像分析 27
3-4-7 細胞粒徑分析 28
3-4-8 細胞SEM攝像 28
3-4-9 生物相容性檢測 29
3-5 統計方法 31
3-5-1 頻率分析 31
3-5-2 顯著性檢定 31
第四章 結果與討論 32
4-1 磁定量技術驗證 32
4-1-1 細胞磁泳試驗 32
4-1-2 磁定量技術-灰階影像分析 37
4-1-3 比色定量分析 42
4-2 磁細胞經表面處理之定性定量分析 44
4-2-1 以磁泳技術分析細胞膜性質變化 44
4-2-2 經表面處理之磁細胞鐵含量檢測 51
4-2-3 經表面處理之磁細胞粒徑分析 54
4-2-4 細胞外部包覆蛋白質之形貌探討 56
4-2-5 經表面處理之磁細胞存活度檢測 58
第五章 結論 61
參考文獻 63

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