臺灣博碩士論文加值系統

　　本研究利用微機電系統製程（Micro-Electro-Mechanical-System, MEMS），成功設計製作3D立體微型磁箝，可以操縱奈米尺度的單一DNA分子，達到拉伸單一DNA分子之操縱，以觀察測量DNA分子之物理機械性質。

　　設計製作之3D立體微型磁箝包含：微型電磁鐵、磁性捕捉器、微流管道等元件。利用田口品質設計方法使磁性合金之電鍍最佳化，降低鍍面之粗糙度，並透過生化修飾，將單一DNA分子兩端分別鍵結直徑為2.8 μm與直徑為1.0 μm的超順磁性磁珠，其中一端之磁珠已經硫醇基修飾，另一端則無。兩端分別鍵結磁珠的單一DNA分子樣品經由蠕動式幫浦輸送到微流管道，當DNA分子到達操作區上方，經由電源供應器施予磁性捕捉器直流電流，產生磁場吸引鍵結磁珠之DNA分子至金表面，已經硫醇基修飾之磁珠與金表面產生共價鍵結，關閉磁性捕捉器之電流後，未經修飾硫醇基之磁珠懸浮在工作溶液中，依序分別施加直流電流給微型電磁鐵，產生磁場吸引懸浮之磁珠，達到拉伸DNA分子之多樣性操縱。微型電磁鐵可產生20pN以上的磁力，將單一DNA分子拉伸至它整個全長以上而不會伴隨有明顯的焦耳熱。DNA分子操縱實驗中，也拉伸了鍵結有兩條DNA分子的磁珠，而DNA也呈現出高度的非線性行為。

　　此3-D微型立體磁箝除了單一DNA分子之操縱外，仍有其他生物應用之潛能，可以將此操縱平台應用在不同之有機線材的研究，例如蛋白質或細胞之研究，透過磁珠鍵結細胞與施加磁場，來觀察細胞。應用此3-D微型立體磁箝操縱平台於生命科學與奈米科技領域。

　　This study designed and fabricated 3-D micromachined magnetic tweezers using MEMS（Micro-Electro-Mechanical-System）technology capable of manipulating a single DNA molecule. The key components, include 6 micro-electromagnets, a ring trapper, a fluidic channel and a gold-patterned surface that can be integrated to form a micromachined-based DNA manipulation platform. The micro-electromagnets can exert a magnetic force over 20 pN with little heating to extend a DNA molecule over the whole contour length.

　　One end of the DNA molecule was bound to a thiol-modified magnetic bead with a diameter of 1.0 μm. The other extremity of the DNA molecule was bound to an unmodified magnetic bead with a diameter of 2.8 μm. A syringe pump was used to transport the DNA sample into the micromachined DNA manipulation platform. Then, the magnetic beads tethered to the DNA molecule were attracted to the gold-patterned surface by the ring trapper. The thiol-modified magnetic bead was immobilized on the gold surface due to the Au-S covalent bonds. When the current of the ring trapper was turned off, the bead with non-specific binding was then suspended in the buffer solution. The suspended magnetic bead could be manipulated within the magnetic field generated by micro-electromagnets.

　　In conclusion, this novel DNA manipulation platform was used to observe the physiological behavior of DNA and investigate its physical properties. The proposed approach will offer a new tool for the field of nano-technology, which will make substantial impacts on development a new nano-scale materials and improve our understanding of intriguing structures.

摘要．．．．．．．．．．．．．．．．．．．．．．．．．．．i
Abstract．．．．．．．．．．．．．．．．．．．．．．．．．． iii
誌謝．．．．．．．．．．．．．．．．．．．．．．．．．．．v
目錄．．．．．．．．．．．．．．．．．．．．．．．．．．．vii
符號說明．．．．．．．．．．．．．．．．．．．．．．．．．x
表目錄．．．．．．．．．．．．．．．．．．．．．．．．．．xi
圖 目 錄．．．．．．．．．．．．．．．．．．．．．．．．． xii

第一章 緒論．．．．．．．．．．．．．．．．．．．．．．．．1
1-2 研究動機及目的．．．．．．．．．．．．．．．．．．．．．2
1-3 文獻回顧．．．．．．．．．．．．．．．．．．．．．．．．3
1-3.1 DNA操縱技術．．．．．．．．．．．．．．．．．．．．．4
1-3.2 傳統磁箝．．．．．．．．．．．．．．．．．．．．．．．12
1-3.3 微型磁箝．．．．．．．．．．．．．．．．．．．．．．．14
1-5 論文架構論述．．．．．．．．．．．．．．．．．．．．．．18

第二章 設計原理．．．．．．．．．．．．．．．．．．．．．． 20
2-1 微磁珠操縱原理．．．．．．．．．．．．．．．．．．．．．20
2-1.1 磁箝理論．．．．．．．．．．．．．．．．．．．．．．．20
2-1.2 微型磁珠之操縱原理．．．．．．．．．．．．．．．．．． 22
2-2 3-D立體微型磁箝設計與操縱原理．．．．．．．．．．．．． 24
2-2.1 3-D立體微型磁箝之設計原理．．．．．．．．．．．．．．． 24
2-2.2 微型電磁鐵之元件設計．．．．．．．．．．．．．．．．． 26

第三章 實驗方法與材料準備．．．．．．．．．．．．．．．．． 29
3-1 微機電製程技術之介紹．．．．．．．．．．．．．．．．．．29
3-1.1 光罩設計與製作．．．．．．．．．．．．．．．．．．．． 29
3-1.2 晶片清潔．．．．．．．．．．．．．．．．．．．．．．．30
3-1.3 金屬薄膜沈積．．．．．．．．．．．．．．．．．．．．．31
3-1.4 微電鍍技術．．．．．．．．．．．．．．．．．．．．．．32
3-1.5 微影製程技術．．．．．．．．．．．．．．．．．．．．．33
3-2 3-D立體微型磁箝之製作．．．．．．．．．．．．．．．．． 36
3-2.1 晶片清潔．．．．．．．．．．．．．．．．．．．．．．．38
3-2.2 金屬薄膜沉積．．．．．．．．．．．．．．．．．．．．．38
3-2.3 第一層光罩：定義底層線圈與磁性捕捉器．．．．．．．．．38
3-2.4 第二層光罩：塗佈電絕緣層及定義底部線圈與頂部之接孔．．43
3-2.5 第三層光罩：定義磁性合金區域與及其電鍍．．．．．．．．44
3-2.6 第四層光罩：定義連接底層線圈與頂層線圈之接孔．．．．．50
3-2.7 第五層光罩：定義塗佈電絕緣層．．．．．．．．．．．．．51
3-2.8 第六層光罩：定義頂層線圈．．．．．．．．．．．．．．．51
3-2.9 第七層光罩：塗佈電絕緣層．．．．．．．．．．．．．．．52
3-2.10 第八層光罩：定義操作區之金區域．．．．．．．．．．． 52
3-3 生物材料之準備．．．．．．．．．．．．．．．．．．．．．52
3-4 實驗設備架設．．．．．．．．．．．．．．．．．．．．．．52

第四章 結果與討論．．．．．．．．．．．．．．．．．．．．． 52
4-1 3-D立體微型磁箝特性分析．．．．．．．．．．．．．．．．52
4-1.1 磁性合金特性分析．．．．．．．．．．．．．．．．．．．52
4-1.2 微型電磁鐵之測試結果．．．．．．．．．．．．．．．．．52
4-1.3 微型電磁鐵之磁力校正．．．．．．．．．．．．．．．．．52
4-2 3-D立體微型磁箝操縱單一DNA分子．．．．．．．．．．．．52
4-2.1 DNA拉伸試驗．．．．．．．．．．．．．．．．．．．．．52
4-2.2 DNA之多樣性操縱．．．．．．．．．．．．．．．．．．．52

第五章 結論與未來展望．．．．．．．．．．．．．．．．．．．52
5-1 結論．．．．．．．．．．．．．．．．．．．．．．．．．．52
5-2 未來展望．．．．．．．．．．．．．．．．．．．．．．．．52
參考文獻．．．．．．．．．．．．．．．．．．．．．．．．． 52

自 述．．．．．．．．．．．．．．．．．．．．．．．．．．．52

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