本論文提出一人工協助生殖之生醫晶片，透過介電泳力與微流道捕捉ICR老鼠的卵母細胞(Oocyte)與精子細胞(Sperm)之生醫晶片，為降低定位時透過接觸力對卵母細胞本身所造成的影響與破壞，本研究將使用非接觸式介電泳力的方式達到卵母細胞的捕捉與體外受精。藉由利用正介電泳力捕捉與定位卵母細胞，以及同時捕捉具有活動力的精子於卵母細胞周圍，來增加卵母細胞周圍的精子濃度與精子總數，對於患有少精症(Oligozoospermia)或是精子活動力較差的男性患者而言，藉由此介電泳力捕捉的方法亦可增加其體外受精的機率，稱之為體外受精技術 (In Vitro Fertilization, IVF)。
本文所提出的介電泳現象，主要是透過一非均勻結構來產生非均勻交流電場，再根據卵母細胞、精子細胞與培養液間之介電係數與導電度的差異，使卵母細胞與精子在非均勻電場受誘導而產生不同偶極矩，使其產生正介電泳效應，而卵母細胞與精子受到正介電泳力的影響會往電場密度較高的地方移動，達到捕捉卵母細胞與精子的效果。此介電泳微流道晶片可於低精子總數(3,000隻)的情況下，提高體外受精率至17.2±7.5 %，比傳統液珠式體外受精的方法提升約21.1 %；且胚胎可正常發育至囊胚期(20.59 %)。

The oviduct is imitated with a biochip system for microfluidic dielectrophoresis (DEP). We show the trapping of an ICR mouse oocyte using the insulator structure of the microchannel in our dielectrophoresis system. To decrease the impact and destruction of the oocyte and the sperm, we adopted a positive dielectrophoretic force to manipulate the oocyte. To imitate a fertilization in vitro (IVF), the oocyte became positioned through the DEP force in the microfluidic channel; then we trapped many more sperm near the oocyte with a positive dielectrophoretic force to enhance the probability of natural fertilization. The oocyte was hence able to become positioned in the micro-channel. At the same time, the sperm swam in the same direction so that many more sperm were trapped near the oocyte to enhance the probability of natural fertilization. Primarily through a non-uniform electric field, the dielectrophoretic force induced the distribution of the electric field.
According to the differences between the relative permittivity and conductivity of the particle and solutions, a difference of dipole moment formed in the electric field. With varied polarization characteristics of the ability of the particles and the buffer solution to generate the positive or negative dielectrophoresis effect, the particles were affected by the DEP force to move to the region with a large or small density of electric field to achieve the goal of oocyte positioning. The positive dielectrophoretic response of an oocyte was exhibited with applied voltage 10 Vpp and frequency 1 MHz; the oocyte was affected by the positive DEP force to move to the region with a large density of electric field. To understand where the positions with a large and small distribution of the electric field in the microchannel, we used commercial numerical software (CFDRC-ACE+) to calculate the location of a large electric field. The pattern of the insulating structure was fabricated with SU8-3050 to generate a non-uniform electric field to trap the oocyte with positive dielectrophoresis. The result of our experiment indicated that a positive DEP served to drive the position of the oocyte and the sperm and to natural fertilization. This DEP microfluidic chip could improve the fertility rate from 14.2±7.5 % to 17.2±7.5 % at the low total sperm number.

致謝 iii
中文摘要 iv
Abstract v
目錄 vi
第一章 緒論 1
1.1 前言與研究背景 1
1.2 研究動機與目的 2
第二章 文獻回顧 8
第三章 介電泳理論 32
3.1 介電泳現象 32
3.2 Clausius-Mossotti Factor 35
第四章 實驗架構與方法 41
4.1 晶片設計 41
4.2 電腦數值模擬 44
4.3 晶片製程 53
4.3.1 電極晶片製作 54
4.3.2 SU8-3050微流道結構製作 59
4.3.3 PDMS翻模 61
4.3.4 氧電漿晶片接合 62
4.4 實驗儀器與系統架構 63
4.4.1 實驗儀器 63
4.4.2 系統架構 66
4.5 實驗樣本處理 66
第五章 研究方法 70
5.1 介電泳緩衝液對精子活動力之影響 70
5.2 介電泳緩衝液對體外受精之影響 73
5.3 介電泳微流體晶片體外受精實驗方法 75
5.4 傳統體外受精實驗方法 79
5.5 電場激活卵母細胞孤雌分裂實驗方法 81
第六章 實驗結果與問題討論 84
6.1 實驗結果 84
6.1.1 介電泳緩衝液對精子活動力影響實驗結果 84
6.1.2 介電泳緩衝液對體外受精影響實驗結果 86
6.1.3 介電泳微流體晶片體外受精實驗結果 91
6.1.4 傳統體外受精實驗結果 99
6.1.5 電場激活卵母細胞孤雌分裂實驗結果 100
6.2 問題與討論 102
第七章 結論 110
第八章 未來計畫 111
第九章 參考文獻 112

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