研究生: |
林彥青 Yen-Chin Lin |
---|---|
論文名稱: |
細菌視紫質生物光電晶片二維掃描系統與訊號探討 2D scanning systems and signal study of bacteriorhodopsin photoelectric chips |
指導教授: |
陳秀美
Hsiu-Mei Chen |
口試委員: |
王鐘毅
Chung-Yih Wang 何明樺 Ming-Hua Ho |
學位類別: |
碩士 Master |
系所名稱: |
工程學院 - 化學工程系 Department of Chemical Engineering |
論文出版年: | 2015 |
畢業學年度: | 103 |
語文別: | 中文 |
論文頁數: | 107 |
中文關鍵詞: | 細菌視紫質 、紫膜 |
外文關鍵詞: | Bacteriorhodopsin, purple membrane |
相關次數: | 點閱:465 下載:2 |
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Halobacterium salinarum 古生嗜鹽菌紫色細胞膜(purple membrane, PM)內含有一單方向光驅動質子泵:細菌視紫質(bacteriorhodopsin, BR),其受脈衝式及連續式雷射激發後分別會產生極快B1B2及微分D1D2光電流響應。本研究利用LabVIEW自動化量測系統,針對BR受光激發後所產生之兩種光電流響應進行訊號快速擷取。研究分為兩部份:第一部份以B1B2光電流訊號之量測來分析PM膜固定化於ITO導電基材上之定向性,結果顯示在PM固定化架橋中添加氧化石墨烯,並對晶片再施以剪應流清洗,可使PM膜排列接近同向且趨近單層,而使得B1光電流訊號大幅增加,同時B2訊號強度減少。接著以微陣列方式掃描PM晶片,進一步證實上述論點。第二部份以D1D2微分光電流分析為量測參數,並利用受檢物可遮蔽入射光而導致PM晶片光電流訊號下降之特性,製作陣列式PM免疫光電感測晶片。首先在PM晶片上分別固定化avidin與NeutrAvidin,接著與高濃度E. coli 反應後,晶片光電流強度分別下降57 %及23 %;接著分別以avidin與NeutrAvidin先固定E. coli抗體於PM晶片上,之後再進行相同菌濃度檢測,晶片光電流訊號則分別下降40 %與50 %,顯示NeutrAvidin對E. coli有較低的非特異性吸附,並以之固定化抗體的晶片對E. coli有較佳檢測靈敏度。同樣以LabVIEW系統掃描陣列式PM免疫光電晶片,可同步且快速地檢測數種菌株。
The purple membrane (PM) of Halobacterium salinarum contains an unidirectional light-driven proton pump, bacteriorhodopsin (BR), which generates a fast (B1B2 ) and a slow (D1D2) photocurrent response upon excitation by a pulse and CW laser, respectively. This research aimed to collect and analyze those two photocurrent responses through a home-built LabVIEW-controlled automatic data acquisition system. This thesis contains two parts. First, the immobilization orientation of b-PM patches on ITO electrodes was investigated by analyzing the B1B2 photocurrent responses. A PM monolayer with nearly uniform orientation could be achieved by a fabrication process with addition of graphene oxide in the linker for PM immobilization and with a followed-up washing over the PM-fabricated chip using a shear flow. This resulted in significant enhance of B1 signals as well as reduction of B2 signals. Microarray-scanning of the as-prepared PM chips also yielded the same supportive observation. Secondly, a PM-based immuno- photoelectric array chip was developed with the D1D2 differentiation photocurrent as the measuring parameter as well as with the fact that PM photocurrents decline when the incident light is blocked by analytes. The detection of Escherichia coli using avidin-bound PM chips with and without prior immobilization of E. coli antibodies resulted in 40% and 57% photocurrent reductions, respectively. On the other hand, 50% and 23% photocurrent reductions were observed when NeutrAvidin-bound PM chips were used to detect E. coli with and without prior immobilization of E. coli antibodies, respectively. Therefore, NeutrAvidin had a lower nonspecific adsorption toward E. coli than avidin, and the prepared antibody-bound PM chips with NeutrAvidin had better detection sensitivity. Finally, simultaneous and fast detection of different organisms was demonstrated by using a LabVIEW-controlled array scanning system.
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陳逸航,"定向性細菌視紫質晶片之光電與二倍頻響應探討",國立台灣科技大學化學工程研究所碩士論文,2010
黃柏竣,"非特異性作用對親和吸附法製備細菌視紫質光電晶片之影響",國立台灣科技大學化學工程研究所碩士論文,2012
林聖為,"利用自排性單分子膜與生物親和吸附增進PM晶片的製程",國立台灣科技大學化學工程研究所碩士論文,2012
林承德,"氧化石墨烯濃度對以親和吸附作用製備細菌視紫質光電晶片之影響",國立台灣科技大學化學工程研究所碩士論文,2013a
林其融,"AFM探討分別以結合氧化石墨烯之親和吸附與共價鍵結作用固定化之紫膜",國立台灣科技大學化學工程研究所碩士論文,2013b
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