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研究生: 陳郁鴻
Yu-Hung Chen
論文名稱: 光學三角測量方法量測家蠶肌肉振動與脈搏變化之研究
The Measurement of Muscle Vibration and Pulse Wave of Bombyx mori Larva Based on Optical Triangulation Method
指導教授: 黃榮南
Rong-Nan Huang
口試委員:
學位類別: 碩士
Master
系所名稱: 生醫理工學院 - 生命科學系
Department of Life Science
畢業學年度: 93
語文別: 中文
論文頁數: 104
中文關鍵詞: 光學三角測量肌肉收縮頻譜
外文關鍵詞: optical triangulation, CMOS image sensor, silkworm, spectrum
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  • 本論文中,我們利用光學三角測量法,成功地利用CMOS sensor、雷射光及MATLAB軟體分析等工具,針對我們所選用的家蠶動物模式,量測其肌肉收縮變化的型態模式,將肉眼不易辨識的動態變化,以精密度達10 μm,數量化細微的物點位移,分別得到各種不同刺激(20% Formalin、4% Acetic acid、10% Formic acid、10% Acetylcholine)的光點變動與時間的關係圖以及頻譜圖。以微注射方式,將上述藥劑各207 nl注射於家蠶背部第七體節後,量測其肌肉收縮之變化,並將波形變動轉換成頻率(或頻譜圖)。結果顯示以空針扎刺之物理傷害與注射PBS(pH 6.5),並未對家蠶幼蟲造成嚴重而持續性傷害,僅於低頻出現amplitude為0.1115與0.1673之波峰;而Acetic acid、Formic acid和Acetylcholine之處理,則較空針扎刺與注射PBS引起之振動有微幅差異,其頻譜低頻最大amplitude分別為0.2380、0.2612和0.2104之波峰,但皆為短暫的振動;而波形之最大振幅,於Acetic acid為2.7 pixel,另兩組(Formic acid和Acetylcholine)跟空針扎刺與PBS皆約1.5 pixel。但Formalin處理引起之蟲體肌肉則有相當明顯的變化,不論從處理前後的波形變化或頻譜的差異性來判讀,振幅最大可達4.5 pixel,低頻amplitude最大為0.8644,顯示Formalin對家蠶幼蟲生理上造成極嚴重的刺激,而關於測得的頻率及振幅與動物疼痛之關聯性與應用,仍須深入之研究。


    This thesis employed the optical triangulation measurement system (including CMOS image sensor, laser light and MATLAB software) to analyze muscle vibration patterns by using silkworm as an animal model. By using this system, the dynamic change of muscle contraction was quantified with 10 μm precision that is unable to be distinguished with naked eye. Each silkworms were nanoinjected with 207 nl of 20% formalin, 4% acetic acid, 10% formic acid or 10% acetylcholine into the 7th segment and muscle vibration patterns were recorded. The acquired vibration information was then transformed into time-position and frequency-amplitude profile. The results showed that puncture and PBS (pH 6.5) injection did not induce significant contraction in silkworm''s muscle. The amplitudes of peaks are 0.1115 and 0.1673 respectively in low frequency. Either acetic acid, formic acid or acetylcholine injection induces short-term vibrations, but distinguishable amplitude change from that of puncture and PBS injection. The amplitudes of the peaks for acetic acid, formic acid or acetylcholine injection are 0.2380, 0.2612 and 0.2104 respectively in low frequency. The amplitude for acetic acid is 2.7 pixel, and for formic acid and acetylcholine are 1.5 pixel, same as that puncture and PBS injection. In contrast, formalin injection induces significant changes of waveform in both amplitude and spectrum. The largest amplitude of waveform is 4.5 pixel and peak in 0.8644 in low frequency. In conclusion, these studies demonstrated the applicability of triangulation measurement system for the physiology of silkworm larva. The relation and application between the frequency, the amplitude and pain of animals deserves further investigation.

    第一章 緒論 Ⅰ. 研究動機與目的 ……………………………………………1 Ⅱ. 家蠶簡介 ……………………………………………………1 Ⅲ. 肌肉收縮 ……………………………………………………4 Ⅳ. 血液及其循環 ………………………………………………7 Ⅴ. 光學三角測量法 ……………………………………………8 Ⅵ. CMOS image sensor ………………………………………10 Ⅶ. MATLAB ……………………………………………………13 Ⅷ. OriginPro …………………………………………………14 第二章 材料與方法 Ⅰ. 研究方法 壹. 數據取得 …………………………………………………15 1. 光學的三角測量法 ………………………………………15 2. 固定 ………………………………………………………19 3. 定義參考點 ………………………………………………19 4. 注射位置 …………………………………………………19 5. 注射劑量 …………………………………………………20 6. 量測系統架構 ……………………………………………21 7. 系統程式控制 ……………………………………………23 貳. 影像分析 …………………………………………………24 1. 轉檔 ………………………………………………………24 2. 亮點閥的設定 ……………………………………………24 3. 質心位置計算法 …………………………………………26 4. 繪圖分析 …………………………………………………27 Ⅱ. 實驗材料 1. 家蠶 ………………………………………………………28 2. 試劑 ………………………………………………………29 第三章 結果 Ⅰ. 三角測量裝置之校正……………………………………31 1. 雷射光參考點穩定度分析 ………………………………31 2. 系統穩定度及校正 ………………………………………32 Ⅱ. 實測結果 …………………………………………………33 1. 五齡家蠶幼蟲經空針扎刺 ………………………………34 2. 五齡家蠶幼蟲注射PBS(pH 6.5)207 nl ………………34 3. 五齡家蠶幼蟲注射20% Formalin 207 nl ………………35 4. 五齡家蠶幼蟲注射4% Acetic acid 207 nl ……………36 5. 五齡家蠶幼蟲注射10% Formic acid 207 nl ……………37 6. 五齡家蠶幼蟲注射10% Acetylcholine 207 nl …………37 Ⅲ. 雷射光參考點變動範圍與時間關係 ………………………38 Ⅳ. 低頻Amplitude與時間關係 ………………………………38 第四章 討論 ……………………………………………………39 第五章 結論 ……………………………………………………43 參考文獻 ……………………………………………………………44 圖表 …………………………………………………………………47 附錄 …………………………………………………………………78

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