臺灣博碩士論文加值系統

本研究旨在開發一種能同時檢測NSE和CYFRA21-1肺癌腫瘤標誌物的微感測晶片。惡性腫瘤是台灣國人十大死因之首，其中肺癌病例逐年增加，且早期症狀不明顯，導致大部分患者在晚期才被診斷出來。因此，早期發現和治療對於提高患者的生存率至關重要。本論文提出了一種基於延伸式閘極場效電晶體(EGFET)的生物感測晶片，並選用高介電常數材料如HfO2、TiO2與Al2O3作為閘極氧化層，以提升感測靈敏度和準確度。
本研究首先介紹了矽基板非等向濕式蝕刻原理及EGFET的工作機理，並詳細說明了高介電常數材料的選擇和應用。在實驗部分，通過KOH濕式蝕刻技術製備感測晶片，使閘極感測區形成類倒金字塔結構，以增加表面積和感測靈敏度。針對不同的閘極氧化層結構，如HfO2/TiO2和Al2O3，進行了多次實驗比較。
實驗結果顯示，使用ALD製程Al2O3閘極介電層的感測晶片在檢測NSE和CYFRA21-1濃度時，表現出最佳的靈敏度和線性度。在NSE量測中，Al2O3閘極感測晶片有最高81 (V·(ng/ml)-1)，並且在最低濃度0.421 ng/ml時仍能準確檢測。在CYFRA21-1量測中，Al2O3閘極感測晶片有最高314 (V·(ng/ml)-1)，並且在最低濃度0.201 ng/ml時仍能準確檢測。相比之下，傳統SiO2閘極的靈敏度和檢測極限均較低。此外HfO2/TiO2閘極感測晶片同樣展示出高靈敏度和低檢測極限，且在高濃度下維持良好的線性關係。
本研究成功開發出高靈敏度、高專一性的微型生醫感測晶片，能有效檢測肺癌相關腫瘤標誌物NSE和CYFRA21-1。未來，該技術有望應用於早期肺癌診斷，提高臨床檢測效率和患者生存率 。

This study aims to develop a micro-sensing chip capable of simultaneously detecting the lung cancer tumor markers NSE and CYFRA21-1. Lung cancer is the leading cause of death among the top ten causes of death in Taiwan, with the number of cases increasing annually. The early symptoms of lung cancer are not apparent, leading to most patients being diagnosed at a late stage. Therefore, early detection and treatment are crucial for improving patient survival rates. This paper proposes a biosensing chip based on an Extended Gate Field Effect Transistor (EGFET), using high-k materials such as HfO2, TiO2, and Al2O3 as the gate oxide layer to enhance sensing sensitivity and accuracy.
First, the study introduces the principles of anisotropic wet etching of silicon substrates and the working mechanism of EGFET. It then details the selection and application of high-k materials. In the experimental part, the sensing chip is fabricated using KOH wet etching technology, forming an inverted pyramid structure in the gate sensing area to increase surface area and sensing sensitivity. Multiple experimental comparisons were conducted on different gate oxide layer structures, such as HfO2/TiO2 and Al2O3.
The experimental results show that the sensing chip with an Al2O3 gate dielectric layer produced using the ALD process exhibited the best sensitivity and linearity when detecting NSE and CYFRA21-1 concentrations. In NSE measurements, the Al2O3 gate sensing chip achieved the highest sensitivity of 81 (V·(ng/ml)-1), and it could accurately detect the lowest concentration of 0.421 ng/ml. In CYFRA21-1 measurements, the Al2O3 gate sensing chip achieved the highest sensitivity of 314 (V·(ng/ml)-1), and it could accurately detect the lowest concentration of 0.201 ng/ml. In comparison, the sensitivity and detection limit of the traditional SiO2 gate were lower. Additionally, the HfO2/TiO2 gate sensing chip also demonstrated high sensitivity and low detection limits, maintaining good linearity at high concentrations.
This study successfully developed a highly sensitive and specific micro biomedical sensing chip capable of effectively detecting lung cancer-related tumor markers NSE and CYFRA21-1. In the future, this technology is expected to be applied in early lung cancer diagnosis, improving clinical detection efficiency and patient survival rates.

論文審定書 i
致謝 ii
摘要 iii
Abstract iv
目錄 vi
圖目錄 ix
表目錄 xii
第一章 緒論 1
1.1 研究動機 1
1.2 文獻回顧 4
1.2.1 電子冷光免疫分析法 5
1.2.2 酶聯免疫吸附測定 5
1.2.3 離子選擇電極 6
1.2.4 表面電漿共振 7
1.2.5 離子感測場效電晶體 8
1.2.6 延伸式閘極場效電晶體 10
1.3 金氧半場效電晶體的高介電常數材料 12
1.3.1 高介電常數材料的選擇 12
1.4 實驗方法與論文架構 14
1.4.1 高介電常數閘極介電層之延伸式閘極場效電晶體 14
1.4.2 生物感測薄膜製作 14
1.4.3 論文架構 15
第二章 原理介紹 16
2.1 矽基板非等向濕式蝕刻原理 16
2.2 延伸式閘極場效電晶體(EGFET)原理 17
2.2.1 能斯特方程式 17
2.2.2 吸附鍵結模型 18
2.2.3 參考電極 19
2.2.4 延伸式閘極場效電晶體工作原理 21
2.3 電晶體閘極氧化層之材料特性 25
2.3.1 閘極氧化層材料對感測靈敏度的影響 25
2.4 高介電常數材料基礎理論 27
2.4.1 電容與等效氧化層厚度 27
2.4.2 高介電材料缺陷對遲滯曲線方向的影響 28
2.4.3 射頻磁控濺鍍(RF Sputter) 30
2.5 理論偵測極限(LOD) 30
第三章 元件設計與製作 32
3.1 具高靈敏度之微型肺癌腫瘤標誌物感測元件 32
3.1.1 具高靈敏度之微型肺癌腫瘤標誌物感測晶片佈局 33
3.1.2 封裝晶片設計佈局 36
3.2 元件製程整合 38
3.2.1 延伸式閘極場效電晶體與微型氯化銀參考電極製作 38
3.2.2 封裝晶片製作與保與飽和氯化鉀洋菜膠調製 60
3.3 感測晶片封裝 65
3.4 NSE/CYFRA21-1感測薄膜調配 68
第四章 實驗結果與討論 70
4.1 微型氯化銀參考電極與閘極介電層薄膜之晶相 70
4.1.1 二氧化鉿薄膜晶相實驗結果與分析 70
4.1.2 二氧化鈦薄膜晶相實驗結果與分析 72
4.1.3 微型氯化銀參考電極實驗結果分析 74
4.2 延伸式閘極場效電晶體電性分析 76
4.2.1 射頻磁控濺鍍薄膜之電壓電容量測結果 77
4.2.2 延伸式閘極場效電晶體閘極介電層之電容值(C-V)分析 78
4.2.3 延伸式閘極場效電晶體之輸出特性曲線(ID-VDS)分析 81
4.2.4 延伸式閘極場效電晶體之閘極漏電流(IG-VGS)分析 83
4.3 肺癌腫瘤標誌物感測晶片量測與分析 84
4.3.1 NSE量測值與靈敏度結果分析 85
4.4 肺癌腫瘤標記物感測晶片專一性量測與分析 90
4.5 肺癌腫瘤標記物感測晶片之比較 93
第五章 結論與未來展望 94
5.1 結論 94
5.2 未來展望 95
參考文獻 97
圖目錄
圖 1-1 112年台灣十大癌症死因[3] ............................................................................ 1
圖 1-2 112年台灣十大癌症死因死亡率[3] ................................................................ 2
圖 1-3 肺癌種類與發生位置[4].................................................................................. 3
圖 1-4 臨床常用之腫瘤標記組合[5].......................................................................... 3
圖 1-5 離，選擇電極(ISE)示意圖.............................................................................. 7
圖 1-6 金屬氧化物半導體場效電晶體(MOSFET)示意圖[10] ................................. 8
圖 1-7 離，感測場效電晶體(ISFET)結構示意圖 ..................................................... 9
圖 1-8 延伸式閘極感測電晶體(EGFET)元件結構示意圖[10] .............................. 11
圖 1-9 高介電材料介電常數與能帶之關係[16] ..................................................... 13
圖 1-10 高介電材料與矽基板的能障接合高度[14]................................................ 13
圖 2-1矽基板非等性蝕刻示意圖[21] ...................................................................... 16
圖 2-2 吸附鍵結模型示意圖[22].............................................................................. 18
圖 2-3 商用參考電極示意圖[24].............................................................................. 20
圖 2-4 金氧半場效電晶體之IDS-VDS壓電特性曲線圖[28] ................................... 22
圖 2-5 延伸式閘極場效電晶體感測器靈敏度判斷示意圖[28].............................. 24
圖 2-6 可移動金屬離，造成電遲滯曲線的效應[30].............................................. 28
圖 2-7 氧化層內電荷造成電遲滯曲線的效應[30].................................................. 29
圖 3-1 延伸式閘極場效電晶體整合微型氯化銀參考電極示意圖 ........................ 32
圖 3-2光罩#1延伸式閘極生醫感測區域立體結構圖形 ........................................ 34
圖 3-3 光罩#2 離，佈植圖形 .................................................................................. 34
圖 3-4 光罩#3源、汲極金屬接觸窗圖形 ............................................................... 35
圖 3-5 光罩#4金屬電極圖形 ................................................................................... 35
圖 3-6 光罩#5微型氯化銀參考電極氯化圖形 ....................................................... 36
圖 3-7 光罩#6 微型氯化銀參考電極圖形 .............................................................. 36
圖 3-8 光罩#7 封裝晶片正面圖形 .......................................................................... 37
圖 3-9 光罩#8 封裝晶片背面圖形 .......................................................................... 38
圖 3-10 製程流程圖：(a)使用PECVD沉積氧化矽作為立體結構阻擋層 (b)定義蝕刻立體結構圖形(光罩#1)、 (c)使用氫氧化鉀對矽基板進行蝕刻出倒金字塔結構 (d)使用BOE去除阻擋層 (e) 使用PECVD沉積氧化矽作為離，佈植阻擋層 (f)定義離，佈植圖形(光罩#2)、 (g) 使用BOE去除阻擋層。............................................................................ 39
圖 3-11(a)使用各式機台沉積閘極氧化層 (b)定義接觸窗圖形還使用BOE進行蝕刻(光罩#3)、 (c)使用電，槍蒸鍍沉積鉻/金 金屬層 (d)定義電晶體源極汲極與閘極區域(光罩#4) (e) 使用電，槍蒸鍍沉積鈦/銀 金屬層 (f) 定義參考電極氯化區域(光罩#5)，並使用鹽酸(HCl)進行氯化 (g) 定義氯化銀參考電極邊緣圖形(光罩#6)，並蝕刻非參考電極圖形區域鈦/銀 金屬層。 .............................................................. 40
圖 3-12 立體感測區製作流程圖 .............................................................................. 44
圖 3-13 延伸式閘極場效電晶體實體圖 .................................................................. 55
圖 3-14 製程流程圖：(a)使用PECVD成長SiO2作為襯墊層 (b)使用PECVD成長氮化矽以作為濕式蝕刻阻檔層 (c)定義封裝晶片正面圖形(光罩#6)、 (d)定義封裝晶片背面圖形(光罩#7)、 (e)使用BOE對氧化層進行雙面蝕刻 (f)使用KOH對矽基板進行雙面蝕刻。 ....................... 60
圖 3-15 晶片封裝示意圖 .......................................................................................... 65
圖 3-16 封裝還之微型肺癌種流標誌物感測晶片 .................................................. 67
圖 3-17 NSE/CYFRA21-1之生物感測薄膜示意圖 ................................................. 69
圖 4-1 立方晶態二氧化鉿(c-HfO2)晶相資料表 ...................................................... 70
圖 4-2 斜方晶態二氧化鉿(o-HfO2)晶相資料表 ...................................................... 71
圖 4-3 本論文二氧化鉿薄膜XRD圖 ...................................................................... 71
圖 4-4 年方晶態金紅石(r-TiO2) 晶相資料表 ......................................................... 73
圖 4-5 本論文二氧化鈦薄膜XRD圖 ...................................................................... 73
圖 4-6 氯化銀 10000倍下SEM圖 ......................................................................... 75
圖 4-7 微型氯化銀參考電極穩定性量測 ................................................................ 75
圖 4-8 HfO2/TiO2 來通入氧氣版本 C-V曲線 ........................................................ 77
圖 4-9 HfO2/TiO2 通入氧氣版本 C-V曲線 ............................................................ 78
圖 4-10 具SiO2 EGFET之C-V特性曲線圖 .......................................................... 79
圖 4-11 具HfO2/TiO2 EGFET之C-V特性曲線圖 ................................................. 79
圖 4-12 具Al2O3 EGFET之C-V特性曲線圖 ........................................................ 80
圖 4-13 具SiO2 EGFET ID-VDS特性曲線圖 ............................................................ 82
圖 4-14 具HfO2/TiO2 EGFET ID-VDS特性曲線圖 .................................................. 82
圖 4-15 具Al2O3 EGFET ID-VDS特性曲線圖 .......................................................... 83
圖 4-16 不同閘極介電層結構EGFET之IG-VGS特性曲線圖 ............................... 84
圖 4-17 HfO2/TiO2 版本NSE抗原響應值與靈敏度量測特性圖 ........................... 86
圖 4-18 Al2O3版本NSE抗原響應值與靈敏度量測特性圖 .................................... 86
圖 4-19 HfO2/TiO2 版本CYFRA21-1抗原響應值與靈敏度量測特性圖 .............. 88
圖 4-20 Al2O3版本CYFRA21-1抗原響應值與靈敏度量測特性圖....................... 88
圖 4-21 HfO2/TiO2 版本NSE感測晶片專一性量測圖 ........................................... 91
圖 4-22 Al2O3版本NSE感測晶片專一性量測圖 .................................................... 91
圖 4-23 HfO2/TiO2 版本CYFRA21-1感測晶片專一性量測圖 .............................. 92
圖 4-24 Al2O3版本CYFRA21-1感測晶片專一性量測圖....................................... 92
表目錄
表 1-1 肺部癌症指數判定[6]...................................................................................... 4
表 3-1 延伸式閘極場效電晶體元件設計規格表 .................................................... 33
表 3-2晶圓標準清洗製程參數表 ............................................................................... 42
表 3-3 KOH結構蝕刻參數..................................................................................... 43
表 3-4 PECVD製程氧化矽參數 ............................................................................... 46
表 3-5 來通氧氣二氧化鉿參數 ................................................................................ 47
表 3-6 來通氧氣二氧化鈦參數 ................................................................................ 47
表 3-7 通入氧氣二氧化鉿參數 ................................................................................ 48
表 3-8 通入氧氣二氧化鈦參數 ................................................................................ 48
表 3-9 ALD三氧化二鋁製成參數 ............................................................................ 49
表 3-10 Ti-cap 層沉積參數 ....................................................................................... 50
表 3-11 快速熱退火參數 .......................................................................................... 50
表 3-12 金屬接觸窗蝕刻參數 .................................................................................. 51
表 3-13 感測晶片製程Run Card .............................................................................. 56
表 3-14 續表 3-13 ..................................................................................................... 57
表 3-15 續表 3-14 ..................................................................................................... 58
表 3-16 續表 3-15 ..................................................................................................... 59
表 3-17 反應式離，蝕刻之氮化矽蝕刻參數 .......................................................... 62
表 3-18 封裝晶片製程Run Card .............................................................................. 64
表 3-19 肺癌腫瘤標誌物生物感測薄膜溶液條配比例 .......................................... 68
表 4-1 斜方晶態和立方晶態HfO2 2-theta理論值與圖 4-3實驗值比較 ............. 72
表 4-2 年方晶態金紅石(r-TiO2)與圖 4-5實驗值比較 ........................................... 74
表 4-3 微型氯化銀參考電極穩定性量測值 ............................................................ 76
表 4-4 不同閘極介電層結構EGFET之電容最大值 .............................................. 80
表 4-5 具HfO2/TiO2閘極介電層之感測晶片於各NSE濃度量測之VOUT .......... 87
表 4-6 具Al2O3閘極介電層之感測晶片於各NSE濃度量測之VOUT .................. 87
表 4-7 具HfO2/TiO2閘極介電層之感測晶片於各CYFRA21-1濃度量測之VOUT ............................................................................................................. 89
表 4-8 具Al2O3閘極介電層之感測晶片於各CYFRA21-1濃度量測之VOUT ..... 89
表 4-9 NSE微型感測晶片與其他設計比較表 ......................................................... 93
表 4-10 CYFRA21-1微型感測晶片與其他設計比較表.......................................... 93

[1] Y. J. Chen et al., "Proteogenomics of Non-smoking Lung Cancer in East Asia Delineates Molecular Signatures of Pathogenesis and Progression," Cell, vol. 182, no. 1, pp. 226-244 , 2020.
[2] 衛生福利部國民健康署. "肺癌防治." https://www.hpa.gov.tw/Pages/List.aspx?nodeid=4050 (accessed.
[3] 中華民國行政院衛生福利部, "111年國人死因統計結果," 2023. [Online]. Available: https://www.mohw.gov.tw/cp-16-74869-1.html.
[4] 馬偕紀念醫院. "肺癌可分成那幾類型?" https://www.mmh.org.tw/taitam/che_int/index4_1_LC04.html
[5] 陳幸鈴. "腫瘤標記檢查之意義." https://wwwfs.vghks.gov.tw/001/VghksUploadFiles/323/ckfile/5969ed4e-f066-4f45-bd0c-9f25ffab2f8b.htm
[6] 林洋鈺醫師. "12大癌症指數大解析－醫師教你如何正確解讀癌症指數." https://www.tyh.com.tw/health/f38.html
[7] N. Obradović, "Solid-contact ion-selective electrodes with high sensitivity – comparison of potentiometric and coulometric signal transduction," Master’s, Faculty of Science and Engineering, Åbo Akademi University, 2019.
[8] 張端芳、林啟萬, "表面電漿，共振式生物感測器之研發與應用," 科儀新知第二十三卷第五期, 2002.
[9] P. Bergveld, "Thirty years of ISFETOLOGY what happened in the past 30 years and what may happen in the next 30 years," Sens. Actuators B: Chem., vol. 88, no. 1-20, 2003.
[10] 廖思媛, "開發具高靈敏度及高專一性之心肌鈣蛋白 I 感測微系統," 碩士, 電機工程學系研究所, 國立中山大學, 高雄市, 2023. [Online]. Available: https://hdl.handle.net/11296/sx762j
[11] P. Bergveld, "Development of an Ion-Sensitive Solid-State Device for Neurophysiological Measurements," IEEE Transactions on Biomedical Engineering, vol. BME-17, no. 1, pp. 70-71, 1970, doi: 10.1109/TBME.1970.4502688.
[12] J. van der spiegel, I. Lauks, P. Chan, and D. Babic, "The extended gate chemically sensitive field effect transistor as multi-species microprobe," Sensors and Actuators, vol. 4, pp. 291-298, 1983.
[13] G. E. Moore, "Cramming more components onto integrated circuits," Proceedings of the IEEE, vol. 86, no. 1, pp. 82-85, 1998.
[14] J. Robertson, "Band offsets of wide-band-gap oxides and implications for future electronic devices," Journal of Vacuum Science & Technology B: Microelectronics and Nanometer Structures Processing, Measurement, and Phenomena, vol. 18, no. 3, pp. 1785-1791, 2000.
[15] D. A. Neamen, "Semiconductor physics and devices: basic principles. 2003," ed: McGraw-hill.
[16] J. A. Kittl et al., "High-k dielectrics for future generation memory devices (Invited Paper)," Microelectronic Engineering, vol. 86, no. 7-9, pp. 1789-1795, 2009.
[17] 陳冠宏, "高靈敏度及高專一性之肺癌腫瘤標記感測微系統開發," 碩士, 電機工程學系研究所, 國立中山大學, 高雄市, 2022. [Online]. Available: https://hdl.handle.net/11296/d94rec
[18] 林佳慧, "開發具溫度補償讀出電路之心肌鈣蛋白I感測微系統," 碩士, 電機工程學系研究所, 國立中山大學, 高雄市, 2021. [Online]. Available: https://hdl.handle.net/11296/2n5y9v
[19] H. Seidel, L. Csepregi, A. Heuberger, and H. Baumgärtel, "Anisotropic Etching of Crystalline Silicon in Alkaline Solutions: I . Orientation Dependence and Behavior of Passivation Layers," Journal of The Electrochemical Society, vol. 137, no. 11, p. 3612, 1990.
[20] P. J. Hesketh, C. Ju, S. Gowda, E. Zanoria, and S. Danyluk, "Surface Free Energy Model of Silicon Anisotropic Etching," Journal of The Electrochemical Society, vol. 140, no. 4, p. 1080, 1993.
[21] "What is MEMS ?" https://www.memsnet.org/mems/fabrication.html
[22] D. E. Yates, S. Levine, and T. W. Healy, "Site-binding model of the electrical double layer at the oxide/water interface," Journal of the Chemical Society, Faraday Transactions 1: Physical Chemistry in Condensed Phases, vol. 70, pp. 1807-1818, 1974.
[23] D. P. Brezinski, "Kinetic, static and stirring errors of liquid junction reference electrodes," Analyst, vol. 108, no. 1285, pp. 425-442, 1983.
[24] C. LibreTexts. "Reference Electrodes." https://chem.libretexts.org/
[25] K. Shimada, M. Yano, K. Shibatani, Y. Komoto, M. Esashi, and T. Matsuo, "Application of catheter-tip ISFET for continuous in vivo measurement," Medical and Biological Engineering and Computing, vol. 18, pp. 741-745, 1980.
[26] R. Smith and D. Scott, "An integrated sensor for electrochemical measurements," IEEE transactions on biomedical engineering, no. 2, pp. 83-90, 1986.
[27] I.-Y. Huang and R.-S. Huang, "Fabrication and characterization of a new planar solid-state reference electrode for ISFET sensors," Thin solid films, vol. 406, no. 1-2, pp. 255-261, 2002.
[28] 施敏 and 伍國, 半導體元件物理與製作技術. 2013.
[29] K. Suzuki et al., "Design and synthesis of a more highly selective ammonium ionophore than nonactin and its application as an ion-sensing component for an ion-selective electrode," Analytical chemistry, vol. 72, no. 10, pp. 2200-2205, 2000.
[30] D. K. Schroder, Semiconductor material and device characterization. John Wiley & Sons, 2015.
[31] 蕭宏, 半導體製程技術導論. 全華圖書股份有限公司, 2024.
[32] D. A. Armbruster and T. Pry, "Limit of blank, limit of detection and limit of quantitation," The clinical biochemist reviews, vol. 29, no. Suppl 1, p. S49, 2008.
[33] C.-H. Hsieh, C.-H. Huang, J.-H. Lin, L.-S. Yu, and I.-Y. Huang, "Development of an EGFET microsensor with 3D structure for high-specificity cardiac troponin I detection," Journal of Micromechanics and Microengineering, vol. 33, no. 4, p. 045001, 2023.
[34] 許博竣, "具高介電常數閘極介電層之高靈敏度微型氯離，感測晶片研發," 碩士, 電機工程學系研究所, 國立中山大學, 高雄市, 2019. [Online]. Available: https://hdl.handle.net/11296/3u92ft
[35] 蘇冠介, "具積體化氯化銀參考電極之可攜式微型氯離，感測晶片開發," 碩士, 電機工程學系研究所, 國立中山大學, 高雄市, 2017.
[36] C. H. Fu et al., "A low gate leakage current and small equivalent oxide thickness MOSFET with Ti/HfO2 high-k gate dielectric," Microelectronic Engineering, vol. 88, no. 7, pp. 1309-1311, 2011.
[37] 藍偉誌、姚聖恩、鄭博任、余志成, "攪拌方式與蝕刻幾何特徵對 KOH矽蝕刻蝕刻速率及表面粗糙度的影響," 國機械工程學會第二十一屆全國學術研討會論文集, 2004.
[38] S. Cheng, S. Hideshima, S. Kuroiwa, T. Nakanishi, and T. Osaka, "Label-free detection of tumor markers using field effect transistor (FET)-based biosensors for lung cancer diagnosis," Sensors and Actuators B: Chemical, vol. 212, pp. 329-334, 2015.
[39] 陳威佑, "高靈敏度神經元特異烯醇酶微型生醫感測晶片之開發," 碩士, 電機工程學系研究所, 國立中山大學, 高雄市, 2021. [Online]. Available: https://hdl.handle.net/11296/7r8w53
[40] K. Si, S. Cheng, S. Hideshima, S. Kuroiwa, T. Nakanishi, and T. Osaka, "Multianalyte Detection of Cancer Biomarkers in Human Serum Using Label-free Field Effect Transistor Biosensor," Sensors & Materials, vol. 30, 2018.