嗜鹽古生菌Halobacterium salinarum紫膜（purple membrane, PM）內具有一種視黃醛蛋白，被稱為細菌視紫質（bacteriorhodopsin, BR）。其受光激發後會使PM膜內外產生質子梯度差，可用以產生光電流。承襲先前本實驗室利用PM晶片光電流訊號強弱和入射光強度呈正向關係以及微生物會散射光之事實所開發出可針對變種鏈球菌（Streptococcus mutans）進行探測之抗體-PM與核酸適體-PM二種複合晶片，本研究首先探討在晶片製程中各加入glycine填補步驟是否可降低此兩種晶片之非特異性吸附問題。結果發現在未加入glycine填補前，S. mutans抗體-PM複合晶片檢測106 CFU/mL S. mutans時，晶片光電流下降比例可達53%；但同時對相同濃度Escherichia coli與Lactobacillus acidophilus檢測時，晶片之光電流下降百分比也分別有21%與24%；但在製程中加入glycine填補後，對E. coli與L. acidophilus檢測後之晶片光電流下降比例則分別下降至8%與7%，證明可大幅降低晶片的非特異性吸附現象。然而S. mutans核酸適體-PM複合晶片對106 CFU/mL E. coli與L. acidophilus檢測時，加入glycine填補步驟後晶片之光電流下降百分比則由原先的14%與19%僅下降至12%與15%，效果較不明顯。

此外，本研究也設計並製作PM光電晶片雙層即時監測微流道，以使PM晶片在緩慢電解液注流下可即時監測上方微流中S.mutans於培養基注流情況下於基材上形成生物薄膜之情況。首先比較修飾有尾端具有不同官能基自排性單分子膜的ITO玻璃表面上生長生物薄膜之形成情形，發現S. mutans於帶正電且較疏水的基材表面上最容易形成具有三維結構的成熟生物薄膜。其次，在胺基化基材上鍵結抗生物薄膜之胜肽後，發現確實能有效抑制S. mutans生物薄膜之形成，且其表面鍵結量越高，效果越好。因此本實驗證實此雙層PM晶片微流道系統可應用於生物薄膜形成之監測。

Bacteriorhodopsin (BR) is a retinal protein residing in Halobacterium salinarum purple membrane (PM). When illuminated, BR pumps a proton gradient across PM, leading to photocurrent generation. To improve the selectivity of antibody-PM and aptamer-PM composite sensor chips for Streptococcus mutans detection, which were previously developed by using the principles that PM photocurrent correlates linearly with illumination intensity and that bacteria scatter light, this study used glycine to block the residual active moiety of the homobifunctional crosslinker fabricated on PM-coated chips for recognition-element conjugation. For the detection of 106 CFU/mL bacteria, without prior glycine blocking, the antibody-PM chips exhibited not only a 53 % photocurrent reduction on S. mutans detection, but also 21% and 24% reductions for E. coli and L. acidophilus, respectively. On the other hand, with prior glycine blocking, the photocurrent reductions of the chips decreased to 8% and 7% on E. coli and L. acidophilus detections, respectively, indicating selectivity improvement of the antibody-PM chips. The effect of glycine blocking was less pronounced with the aptamer-PM chips because the photocurrent reductions decreased from 14% and 19% to only 12% and 15% for E. coli and L. acidophilus, respectively.

Furthermore, a double-decked microfluidics device was designed and constructed to real-time monitor the formation of S. mutans biofilm on the substrate placed in the upper deck with a PM-coated photoelectric chip mounted in the lower deck. A diluted growth medium and an electrolyte buffer were injected into the upper and lower decks, respectively. The comparison of ITO glass modified with self-assembled monolayers with different terminal groups suggested that the hydrophobic and positively charged surface most easily caused S. mutans to form a mature 3D-strucutred biofilm. Moreover, the anti-biofilm peptides coated on the amine-terminated substrate effectively inhibited biofilm formation, with the inhibition efficiency increasing with their coating amounts. Therefore, the results confirmed the feasibility of real-time monitoring biofilm formation with the developed double-decked microfluidics.

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