臺灣博碩士論文加值系統

本篇論文描述我們實驗室自行架構的雷射掃描共焦式光學顯微鏡，其中在光學系統內包含奈米移動平臺與振鏡兩種掃描，利用振鏡的快速能大範圍掃描獲取影像結合奈米移動平臺局部掃描精準尺寸，我們各自取其優點來大幅增加雷射掃描共焦式光學顯微鏡的使用效率。
由於兩種不同的掃描方式造就彼此影像上有所差異，主要透過相位相關法搭配大津(Otsu)法自動選取最佳二值化閾值與內插法應用來精算平移、旋轉、縮放等參數，得到參數後再將振鏡影像匹配校準至平台影像，結合演算法與人機介面使得本系統在儀器操作上與分析數據和影像校準上更加的人性化與自動化，並用數組數據證實影像差異校準與本系統分析程式的精確度，成功建立一套匹配校準影像程序。

The microscope we use is a laser scanning confocal microscope that combines two scanning systems of nanopositioning stage and galvanometer. Scanning by using the nanopositioning stage is able to take images of high resolution, whereas the galvanometer scanning system allows fast image scanning with a limited resolution. To take advantage of both systems, it is necessary to calibration the images taken by using both scanning systems.
The two different scanning methods cause image divergence in translation, rotation, and scaling, etc. To remove the image divergence, we develop an image calibration procedure by using the Otsu method to acquire the best binarization automatically, and then by utilizing phase correlation with interpolation to calculate the translation, rotation, and scaling parameters for accurately calibrating the galvanometer image to the nanopositioning stage image. We developed a computation process and a user-friendly interface to automatically accomplish the calibration after the images are taken.

中文摘要 i
英文摘要 ii
誌謝 iii
目錄 iv
圖目錄 vi
第一章 緒論 1
1.1 前言 1
1.2 共焦式顯微鏡簡史 2
1.3 研究動機與目的 3
第二章 研究背景與理論基礎 5
2.1 光學成像原理 5
2.1.1 線性非時變系統 5
2.1.2 光學解析度 6
2.1.3 點擴散函數 8
2.1.4 系統成像 9
2.2 共焦式顯微鏡成像原理 10
2.3 傅立葉轉換 15
2.3.1 離散傅立葉轉換 15
2.3.2 快速傅立葉轉換 16
2.4 相位相關法 17
2.4.1 相位相關法平移量計算 18
2.4.2 相位相關法旋轉角度與縮放倍率計算 19
2.5 灰階級量化與二值化 20
2.6 大津演算法Otsu 24
2.7 數位影像處理 26
第三章 實驗系統架構與流程 27
3.1 光學系統與硬體設備 27
3.1.1 雷射光源 28
3.1.2 樣品掃描系統裝置 28
3.1.3 共軛聚焦裝置 29
3.1.4 訊號偵測裝置 30
3.2 控制程式介面 31
3.2.1 粗調平臺控制 31
3.2.2 微調平臺控制 32
3.2.3 平臺掃描系統 33
3.2.4 振鏡掃描系統 35
3.3 系統操作流程 36
3.4 影像映射與校準 37
3.5 程式自動化數據分析 41
第四章 實驗結果與分析 46
4.1 內插法與精確度 46
4.2 顯微校正片測試影像映射與校準 52
4.3 螢光樣品影像映射與校準 59
第五章 結論與未來展望 69
5.1 結果與討論 69
5.2 未來展望 70
參考文獻 71

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