雷射掃瞄共焦光學顯微鏡結合平台與振鏡系統之程式自動化校準影像程序

Abstract

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.