臺灣博碩士論文加值系統

在許多斑馬魚微流體裝置中，麻醉後的斑馬魚幼魚需要被傳送至測試區，而傳送的方式往往為透過機械性質的輸送方法來傳輸斑馬魚幼魚，透過這種傳輸方式，可能會對幼魚造成永久性的損傷，因此，若能透過非侵入式的運送方式輸送斑馬魚幼魚，便能防止幼魚受接觸而受到的損傷；由於視覺與流體力學的刺激容易影響斑馬魚幼魚的游動行為，因此若能利用幼魚本身的天生行為，使其能透過自身的移動而達到實驗的傳
輸目的，便可避免幼魚因強制傳輸而受到的傷害，本實驗框架使用前述的視覺與流體力學刺激控制斑馬魚幼魚，以達到迅速及有效的傳輸目標，為了能提供有效的流體力學刺激，流量會經調整以對斑馬魚幼魚的側線系統 (Lateral line)有所影響，與此同時，視覺刺激由電腦產生持續移動的黑白條紋光流 (Optic flow)，以促使斑馬魚幼魚的視動行為 (Optomotor response)產生，最後透過不同條件搭配的實驗結果，可以觀察到兩種刺激模式的強度都對斑馬魚幼魚的行為有至關重要的影響，以 5 dpf (days-postfertilization)的斑馬魚幼魚來說，在流量 0.1 ml/min 下，並搭配 1 赫茲頻率的光流，其平均傳輸時間為 1.29 秒，相較於只提供流動刺激的傳輸時間，大約只有其三分之
一的時間，此實驗框架可與其他相關為流體裝置做整合，並且可以進一步用於藥物測試，傳輸未麻醉的斑馬魚幼魚至不同方向的測試區。

Most of microfluidics applied to larval zebrafish screening need to transport zebrafish larvae to the designated region of microfluidics. Conventionally, zebrafish larvae is transported by mechanical or manual means, resulting in the paramount damage to larval zebrafish. Therefore, through a noninvasive way, the damage caused by transportation could be eliminated or minimized. Due to visual cues and hydromechanical cues affecting the behavior of zebrafish strongly, we propose an experiment framework that would apply both cues to zebrafish, towards their transportation. In order to provide hydromechanical cues, flow rate would be adjusted to affect the lateral line of larval zebrafish meanwhile to provide visual cues, pattern composed of black and white stripes was produced by the computer animation that drives the optomotor response of zebrafish larvae. From the results, it was observed, both modalities play crucial role in the process of transportation. For 5 days-post fertilization zebrafish larvae, with flow rate of 0.3 ml/min along with the pattern of 1Hz temporal frequency, the average transportation time period is 1.07 This experiment frame work can be used to transport the non-anesthetized larval zebrafish into different section of microfluidics for drug testing.

摘要 I
Extended Abstract II
目錄 V
表目錄 VIII
圖目錄 IX
第1章 緒論 1
1.1 研究背景 1
1.2 斑馬魚 (Danio rerio) 2
1.2.1 斑馬魚動物模型 2
1.2.2 斑馬魚行之趨流性 (Rheotaxis) 4
1.3 斑馬魚微流體裝置之發展 7
1.3.1 斑馬魚胚胎微流體裝置應用 10
1.3.2 斑馬魚幼魚微流體裝置應用 14
1.4 研究動機 17
1.5 研究目標 18
第2章 材料與研究方法 19
2.1 斑馬魚飼養與幼魚準備 19
2.2 微流體裝置的設計與製造 20
2.3 實驗平台架設與影像分析 24
2.4 實驗方式 26
2.4.1 斑馬魚幼魚在視覺與流動刺激下的直線位移實驗 26
2.4.2 流動刺激與視覺刺激對斑馬魚幼魚之影響強度實驗 27
2.4.3 斑馬魚幼魚在流動刺激下受視覺刺激之側向位移實驗 27
2.5 分析方法 28
2.5.1 幼魚移動時間量化分析方式 28
2.5.2 游動之幼魚動能參數分析 29
2.5.3 視覺與流動刺激影響強度分析 30
2.5.4 幼魚側向位移之位置分析 30
2.5.5 統計學分析 31
第3章 結果與討論 32
3.1 斑馬魚幼魚相應於視覺與流動刺激下的行為反應 32
3.2 游動幼魚之動能參數 35
3.3 斑馬魚幼魚對應於生長時間增加之行為變化 39
3.4 流動與視覺刺激對斑馬魚幼魚的影響強度 40
3.5 透過流動刺激與轉向光流傳輸斑馬魚幼魚至不同的測試位置 42
第4章 結論與未來發展 45
4.1 結論 45
4.2 未來發展 46
參考文獻 47

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