細菌培養技術應用在微生物學已經成熟，可是在液體上的過量消耗以及機台的維護上所需費力，在體積大的生物反應器上現代已經越來越微小化在晶片上，我們使用了大腸桿菌定期稀釋以及將生物反應室最小化分成兩部分，來確實執行模擬出宏觀生物反應器，我們設計了微流體晶片生物反應器(microfluidic bioreactor)，這組裝置可執行自動化，長時間培養在nanoliter等級上的生物培養，跟即時的顯微鏡測量計算，因為微流體晶片的高面積體積比，可以有效地將浮游生物及生物膜共同生長，我們期待是有機會來時驗出更多的生物實驗。

Bacterial culture is a basic technique in both fundamental and applied microbiology. The excessive reagent consumption and laborious maintenance of bulk bioreactors for microbial culture have prompted the development of miniaturized on-chip bioreactors. With the minimal choice of two compartments (N = 2) and discrete time, periodic dilution steps, we realize a microfluidic bioreactor that mimics macroscopic serial dilution transfer culture. This device supports automated, long-term microbial cultures with a nanoliter-scale working volume and real-time monitoring of microbial populations at single-cell resolution. Because of the high surface-to-volume ratio, the device also operates as an effective biofilm-flow reactor to support cogrowth of planktonic and biofilm populations. We expect that such devices will open opportunities in many fields of microbiology.

目錄
一、緒論 1
1-1 研究動機 1
1-2 恆化器(chemostat) 3
1-3 微型生物反應室(microbioreactor) 6
1-4 生物膜 (Biofilm) 7
1-5 微流體晶片介紹 8
1-5-1 光罩(Mask) 8
1-5-2 膜仁（Mold） 8
1-5-3 閥門（Valve） 9
1-5-4 蠕動汞浦（peristaltic pump) 10
二、微流體晶片 11
2-1 膜仁製作(Mold) 11
2-2微流體晶片主體製作(Fabrication) 12
2-3 Bioreactor晶片結構(Structure) 14
2-4 Bioreactor稀釋模式(Dilution) 15
2-5 數學理論模型(Math) 16
三、系統架設 17
3-1 Labview晶片控制系統(Control) 19
3-2 溫度控制系統(Temp) 23
3-3 雙螢光數位顯微鏡(Digital Microscope) 25
3-4 Matlab計算(Count) 26
四、大腸桿菌長時間培養 28
4-1 大腸桿菌樣本準備 28
4-2設定以及實驗數據 28
4-3 長時間培養結果 29
五、結論 32
附錄 33
A-1 Bioreactor數學理論分析 33
A-1-1 簡介 33
A-1-2 伴隨著牆壁效應的Bioreactor模型 34
A-1-3 伴隨著牆壁效應Bioreactor模型的解析解 35
A-2 Matlab數學模擬程式碼 39
A-3 Matlab螢光微生物亮度強度計算程式碼 42
A-4 Arduino控制PWM溫度控制程式碼 43
中英專有名詞對照表 45
參考文獻 47






圖目錄
1-1 恆化器示意圖 2
1-2 離散恆化器 4
1-3 16個隔室的封閉迴圈以及大腸桿菌成長曲線 5
1-4 生物膜演化示意圖 6
1-5 光罩圖 7
1-6 膜仁 8
1-7 閥門運作方式 8
1-8 蠕動汞浦 9

2-2 微流體晶片製作流程 12
2-2 晶片設計的局部放大圖 13
2-3 稀釋流程 14

3-1 整體架構 16
3-2 上下層系統架設完整圖 17
3-3 下層多功能檢測系統電路 17
3-4 Labview及時控制顯示面板 19
3-5 管線的配置 20
3-6 溫度控制系統 22
3-7 Arduino控制溫濕度顯示在lcd版 22
3-8 零件裝置圖 23
3-9 雙螢光數位顯微鏡 24
3-10 物竟聚焦在整體圓環上 24
3-11 螢光亮度計算流程圖 26

圖 4-1 微流體管道稀釋 27
圖 4-2 螢光亮度成長曲線 29
圖 4-3 螢光亮度成長曲線 29
圖 4-4 生物膜在微流體晶片內成長示意圖 30
圖 4-5 使用倒立顯微晶觀察到的生物膜成長 30

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