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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 薛景中 | |
dc.contributor.author | Kuo-Jui Chu | en |
dc.contributor.author | 朱國瑞 | zh_TW |
dc.date.accessioned | 2021-06-15T12:56:51Z | - |
dc.date.available | 2017-07-26 | |
dc.date.copyright | 2016-07-26 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2016-07-14 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/50761 | - |
dc.description.abstract | 二次離子質譜儀(SIMS)在表面分析上具有極高的靈敏度。加上近年來簇離子團(cluster ion)濺射技術的發展,二次離子質譜儀能夠產生高質量的離子破片,並分析未經同位素標記的混合樣品。而飛行式二次離子質譜儀(ToF-SIMS)屬於靜態二次離子質譜儀,在分析樣品後不對其造成改變,且能獲得全質譜以利分析。此外,利用高度聚焦的一次離子掃描樣品表面能獲得足夠的空間解析度,而且只需要少量的樣品消耗就能取得高解析度的分子分佈影像。結合微陣列(microarray)能善用上述優點進行高通量的樣品分析。近來由於成本低廉與製備容易,紙質的微陣列是與SIMS分析整合的絕佳選擇。在此研究中則採用玻璃纖維濾紙來製作微陣列,因為無機的玻璃纖維相較於一般的紙質濾紙在高質量區的質譜中有較低的背景質,較適合用來分析。玻璃纖維的濾紙會經octadecyltrichlorosilane (OTS)的自組裝單層膜(self-assemble monolayer)修飾成疏水性的表面後,再使用雷射雕刻機選擇性的製作出親水性的微陣列。接著不同濃度的胜肽(peptide)水溶液以1 μL分別滴在這些親水的圓形區域中,而尚未移除的疏水區域用來區隔每個粒滴。用來獲得二次離子影像的脈衝20 kV C60+聚焦至∼3 μm,離子通量為3.8×109 ion/cm2。大面積馬賽克影像是由700 μm×700 μm每幀256×256像素的影像拼成。經由實驗發現,胜肽分子的二次離子訊號相對強度正比於在水溶液中的濃度,透過不同胜肽分子濃度對二次離子相對強度的數據能繪製校準曲線用來定量混合的胜肽分子。藉由結合玻璃纖維基材的微陣列與影像二次離子質譜儀能高通量且低樣品消耗的分析混合胜肽樣品。 | zh_TW |
dc.description.abstract | Secondary ion mass spectrometry (SIMS) is known as a powerful technique to perform surface analysis with high sensitivity. In recent development of cluster ion sputtering, current SIMS is capable to generate molecular ions of high mass that allow the parallel analysis of molecular mixtures without the need of labeling. With time-of-flight SIMS that operated within the static regime, the specimen is practically unaltered after analysis and the whole m/z pattern (up to a few thousand) can be acquired at the same time. Furthermore, with a focused primary ion that scans over the specimen, high spatial resolution images of molecular distribution on a surface could be obtained with minimal sample consumption. In order to utilize these advantages, a microarray of specimens could be prepared for automatic high throughput analysis of trace molecules. Recently, owing to its low-cost fabrication and ease of modification, paper-based microarray is an excellent candidate to be integrated with the SIMS analysis. In this work, glass microfiber filter paper was used to prepare the microarray because the inorganic glass microfiber is found to yields less background in the high mass (molecular ion) regime than cellulose-based paper. The filter paper was modified with octadecyltrichlorosilane (OTS) self-assemble monolayer (SAM) to form a hydrophobic surface. Hydrophilic microarray was then prepared by using a laser scriber that selectively removed the OTS layer with a designed pattern. 1 μL aqueous drops of various peptides at concentrations between 1.0 and 0.1 mM were then dispensed inside these circular patterns where the OTS SAM was removed while the remaining OTS layer serves as a barrier and separated each drop. Secondary ion images were acquired with a focused (~3 μm beam size) C60+ pulse operated at 20 kV and the primary ion dose was 3.8×109 ion/cm2. Large area mosaic image was obtained by stitching 700×700 μm2 frames of 256×256 pixel. The secondary ion intensities of peptides (e.g. M+Na+) normalized with respect to glass microfiber (SiOH+) were found to be proportional to their concentration in the solution. In other words, calibration curve for each peptide was generated for quantification and the detection limit is in the order of sub-pmol. By combining the paper-based micropad and imaging SIMS, this work demonstrated that the composition of mixed peptides can be determined with high throughput and low sample consumption. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T12:56:51Z (GMT). No. of bitstreams: 1 ntu-104-R02527047-1.pdf: 9001615 bytes, checksum: 662f2c2219cb3a181d82f077bc41a66c (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 口試委員會審定書 #
誌謝 i 中文摘要 iii ABSTRACT iv 目錄 vi 圖目錄 ix 表目錄 xii 第1章 緒論 1 第2章 文獻回顧 4 2.1 質譜儀於解析生物及有機分子之應用 4 2.2 二次離子質譜儀於分析生物及有機分子之應用 9 2.2.1 簇離子與多原子離子應用於二次離子質譜儀之演進 10 2.2.2 簇離子與多原子離子濺射機制 12 2.3 C60+離子源於二次離子質譜儀之應用 14 2.3.1 C60+離子源濺射於SIMS中的優勢 — 二次離子產率提高 14 2.3.2 C60+離子源濺射於SIMS中的優勢 — 低損傷機率 15 2.3.3 C60+離子源濺射於SIMS中的優勢 — 有助影像建立 15 2.4 C60+離子濺射中以基質(matrix)輔助之分析 19 2.5 自組裝單層膜(self-assembled monolayer, SAM) 20 2.5.1 有機矽氧烷自組裝膜 22 2.6 高通量分析(High-throughput analysis) 25 2.6.1 微陣列分析(Microarray) 25 2.6.2 紙基微陣列(paper-based microarray) 27 第3章 實驗 30 3.1 藥品與基材 30 3.2 實驗儀器介紹 31 3.2.1 飛行時間二次離子質譜儀(Time-of-Flight Secondary Ion Mass Spectrometer, ToF-SIMS) 31 3.2.2 掃描式電子顯微鏡(Scanning Electron Microscope) 33 3.2.3 CO2雷射雕刻機(CO2 Laser Scriber) 33 3.3 實驗步驟 35 3.3.1 微陣列製備 35 3.3.2 樣品製備 35 3.3.3 ToF-SIMS分析 36 3.3.4 SEM分析 37 第4章 實驗結果與討論 38 4.1 微陣列製備 38 4.1.1 基材選擇 38 4.1.2 直接觀察微陣列 40 4.1.3 SEM觀察微陣列 41 4.1.4 微陣列的二次離子影像 45 4.2 基材與各胜肽之定性分析 48 4.2.1 基材分析 48 4.2.2 各胜肽(peptide)分子之定性分析 49 4.2.3 小結 57 4.3 各胜肽分子之定量分析 58 4.3.1 高通量之胜肽分子定量分析 58 4.3.2 混合胜肽之定量測試 65 第5章 結論 67 參考文獻 68 | |
dc.language.iso | zh-TW | |
dc.title | 以紙基微陣列與分子型二次離子質譜術進行高通量之
平行檢測與定量 | zh_TW |
dc.title | High-Throughput Parallel Detection and Quantification of Peptides with Paper-based Microarray and Molecular
Time-of-flight Secondary Ion Mass Spectroscopy (ToF-SIMS) | en |
dc.type | Thesis | |
dc.date.schoolyear | 104-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 虞邦英,康佳正 | |
dc.subject.keyword | 二次離子質譜儀,微陣列,高通量分析, | zh_TW |
dc.subject.keyword | Secondary ion mass spectrometry,microarray,high-throughput analysis, | en |
dc.relation.page | 73 | |
dc.identifier.doi | 10.6342/NTU201600246 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2016-07-15 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 材料科學與工程學研究所 | zh_TW |
顯示於系所單位: | 材料科學與工程學系 |
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