根據衛福部資料統計台灣地區育齡期夫妻中不孕之比例約為10~15%，而美國國家疾病管制局(Centers for Disease Control and Prevention)統計數據指出，在美國大約有11％的女性在15~44歲時即面臨懷孕困難，甚至是不孕症。面臨如此高的不孕症比例，民國103年台灣地區的體外受精技術（IVF）成功懷孕率卻只有33％，造成如此低成功率除了精子細胞與卵子細胞的品質以外，其中很重要的一個原因為卵細胞透明帶去除的過程，為了要將卵細胞外層的透明帶(Zona)去除，傳統生物醫學實驗室的作法非常繁雜，利用酵素降解去除透明帶以及培養液浸泡沖洗卵細胞的方法，卵細胞前後至少需經過至少8次的轉移，為了將酵素對卵細胞的傷害降到最低，因此完成一次透明帶去除的過程約耗時30分鐘，而利用取卵針直接吸取卵細胞且頻繁的移動卵細胞也很容易對卵細胞產生傷害。
因此本研究開發一種三維結構微流體晶片，能夠以更溫柔的方式完成透明帶去除過程並且固定卵細胞位置並置換周圍流體以簡化透明帶去除的過程，將原先需耗時30分鐘的去除透明帶過程簡化至只約需4分鐘的過程，且能夠在單一晶片上同時讓多顆卵細胞進行透明帶去除，除此之外利用本研究開發的晶片能夠方便追蹤觀察每一顆卵細胞健康情況，本研究成功利用高分子材料(PDMS)，除了生物相容性高，適合於醫學檢測，且其價格較低，可大量生產製造，將有效降低以往體外受精技術卵細胞製備所需時間以及成本。

According to the statistics of Center for Disease Control and Prevention, there are about 11% of women at 15–44 year of age in the United States encountering difficulty for pregnancy regardless of marital status. Ought to the high rate of infertility, artificial insemination becomes more important to assist on the infertility issues. IVF increases the rate of pregnancy however the success rate is only 35%. One of key points to this low successful rate was stemmed from the protection shell of oocytes retarding sperm self-inseminations. Therefore, cumulus mass and zona pellucid removal using enzyme medium (hyaluronidase and trypsin) was proposed by Yujiroh Kamiguchi [2] to assist sperms’ insemination. Although widely used, however there consists of a serious concern regarding that the oocyte must be shifted to different medium frequently and easily to get severe hurt during the shifting processes. To solve this problem, a 3D microfluidic device is designed in this study and demonstrated the utility on mice oocyte. The device can accommodate oocyte at a specific position and allow pipetting/exchanging enzyme medium/buffers into the position directly without moving oocyte. In this way, cumulus mass and zona pellucid can be detached from oocytes safely and gently at the same time before oocytes used in IVF.

目 錄
摘 要 i
致 謝 iii
圖目錄 vii
表目錄 x
第一章 緒論 1
1.1 研究背景 1
1.1.1微機電系統 1
1.1.2微流體生醫晶片 2
1.1.3不孕症數據統計 3
1.2 研究動機 5
第二章 文獻探討 8
2.1細胞捕捉與定位 8
2.2微流體交換系統 11
2.3 研究目的 14
第三章 實驗設計與規劃 15
3.1 實驗原理 15
3.2 晶片設計 17
3.2.1微流道結構 17
3.3 實驗藥品與儀器 18
3.3.1 實驗藥品 18
3.3.2 實驗儀器 20
3.5 實驗步驟 22
3.5.1 微流道製作 22
3.5.2孔洞薄膜製作 26
3.6 實驗流程 27
3.6.1 母倉鼠誘導排卵 27
3.6.2 透明帶去除 28
第四章 結果與討論 30
4.1 第一代晶片設計及實驗 30
4.1.1微流道測試 30
4.1.2薄膜製程結果 31
4.1.3稀釋濃度測試 32
4.1.4鼠卵測試與培養 34
4.2 第二代晶片設計與實驗 35
4.2.1 複合形薄膜製程結果 36
4.2.2 流速測試 39
4.2.3 溫度測試 40
4.2.4 單顆卵細胞測試 43
4.2.5 多顆卵細胞測試 44
4.2.6 與傳統去除透明帶方法受精成功率比較 46
第五章 結論 49
第六章 未來發展方向 50
第七章 參考文獻 51

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