過去許多學者研發不同觀察分析或者訓練果蠅行為的系統，這些觀測分析以及行為訓練系統各自有各自的用途以及特色，然而過去本實驗室開發一套研究果蠅行為的即時影像分析與雷射刺激之系統，此系統能夠即時分析及記錄果蠅社交行為並搭配外部的雷射刺激源應用在果蠅的Chlamydomonas reinhardtii channelrhodopsin (ChR2)及Natronomonas pharaonis halorhodopsin (NpHR)相關的實驗上。本論文研發自動化即時分析與溫度調控果蠅社交行為之系統，此系統不只研究由雷射控制的ChR2與NpHR實驗上，還結合新的外部刺激源，而此刺激源是由高溫(31℃)調控Transient Receptor Potential (dTrpA1) 與 shibire(shits1)的相關實驗，若能夠如此應用兩種外部刺激源可以了解果蠅腦神經的網路溝通結構圖，知道神經網路上下游是如何進行溝通。
　　本論文之系統是由本實驗室之前所開發的”果蠅行為的即時影像分析與雷射刺激之系統”為基準進行改良及開發，新系統延續之前即時追跡及分析果蠅行為搭配外在雷射刺激源，而新系統增加溫度調控的外部刺激源，此刺激源使果蠅觀測盒(Arena)部分達到高溫(31℃)，藉以達到激發或抑制果蠅行為之效果。一開始是設計一個溫度調控箱將系統罩住進行加熱以維持恆溫的系統，但此系統有一些缺點(ex:加熱時間太久、無法降溫)，所以我們改變加熱源的加熱方式，以鹵素燈搭配長通濾光鏡進行加熱，並改善溫度調控箱的缺點。這套系統能夠提供給實驗者從基本的資料分析到搭配不同外部刺激源作為訓練，以達到激發或抑制果蠅的特定行為表現，使得研究人員在實驗的設計上擁有更大的自由度以及操作上更豐富的系統功能。

In past several researchers developed different kind of analyzing and training systems for Drosophila social behavior, different systems had their own advantages and characteristics. However our laboratory develop “Camera-Base Real-time Analysis with Laser Stimulated for Behavior Study of Drosophila“, this system not only can real-time analysis, but also activated Chlamydomonas reinhardtii channelrhodopsin (ChR2) and inhibited Natronomonas pharaonis halorhodopsin (NpHR) by two kind of Laser. In this thesis develop “Automated Real-time Analysis and Temperature Control System for Drosophila Social Behavior”, this system not only can research ChR2 and NpHR experiment by laser, but also can research Transient Receptor Potential (dTrpA1) and shibire (shits1) experiment at higher temperature (31℃). By using two kind of external stimulus, we can understand the neuron networks of Drosophila brain.
In this thesis, the new system is developed by “Camera-Base Real-time Analysis with Laser Stimulated for Behavior Study of Drosophila“, the system add new external stimulus by temperature control , this stimulus let Arena at 31℃, by this stimulus can activate dTrpA1 and inhibit shibire. First we design a temperature control box covered system, by this way the system can heat and maintain at 31℃, but this system has several disadvantages (ex: waste long heating time, can’t cooling). So we design new heating stimulus, it make by halogen lamp and long pass filter.

致謝 I
摘要 II
ABSTRACT III
目錄 V
圖表目錄 VI
第一章　緒論 1
1.1 研究背景 1
1.2 文獻回顧 2
1.2.1 果蠅影像處理 2
1.2.2 果蠅學習與記憶平台 3
1.3研究動機 5
1.4論文計劃書架構 5
第二章　系統平台的架構與設計 16
2.1 硬體架構 (HARDWARE) 16
2.1.1分析系統模組 16
2.1.2 CCD Camera 17
2.1.3果蠅行為觀測盒與觀測盒置放機構 18
2.1.4白光LED模組 19
2.1.5雷射掃描模組 20
2.1.6溫度調控模組 20
2.2 軟體架構 (SOFTWARE) 21
2.2.1 LabVIEW 圖形化程式 21
2.2.2人機介面(GUI, Graphical User Interface) 22
2.2.3果蠅追蹤雷射訓練程式 23
第三章　實驗設計與初步實驗結果 42
3.1實驗一 確認本系統是否維持在工作溫度 42
3.2實驗二 利用本系統達到抑制果蠅行為 42
3.3實驗三 不同溫度下對於公果蠅的求偶行為影響 43
第四章　新一代系統設計與實驗結果 53
4.1上一代系統缺點及改善方式 53
4.2硬體架構設計 53
4.3軟體參數修正 54
4.4實驗一 ARENA溫度變化 55
4.5 實驗二 新一代溫度抑制果蠅行為 55
4.6 實驗三 新一代系統溫度激發果蠅行為 56
4.7 實驗四 高溫下果蠅求偶行為表現 57
第五章　結論 66
第六章　未來工作 68
參考文獻 69

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