非線性光學顯微鏡的建立與其工業上之應用

Abstract

在偏振片中聚乙烯醇（PVA）層是主要產生偏振功能的光學元件，在一般偏振片的製造過程中聚乙烯醇層的碘濃度無法被即時監測，需要通過切片放到電子顯微鏡下檢查才能知道碘濃度分布。但是這樣的檢測是有侵入性、破壞性和非即時的。在這次的研究中，我們提出了快速與非侵入性的樣品掃描非線性光學顯微鏡。藉由樣品掃描非線性光學顯微鏡，我們可以區分聚乙烯醇層中碘濃度的分布是否均勻。利用軸向的深度掃瞄，可以得知碘濃度分布均勻會有一個吸收峰，碘濃度分布不均勻則會有兩個吸收峰。另一方面，為了提高非線性光學顯微鏡的成像速度，我們利用的雙軸Galvano鏡架設出光束掃瞄非線性光學顯微鏡。在光束掃瞄非線性光學顯微鏡中，利用了Resolution Chart確定解析度與圖像大小。得到的最佳的解析度是0.87μm和250μm*250μm的影像大小。最後我們利用不同條件的雷射光測量出葉肉細胞與葉綠體的雙光子螢光影像。

Polyvinyl alcohol (PVA) layer is the key layer in optical polarizer. During its fabrication, the PVA concentration cannot monitored in real time. It should be accomplished by slicing or biopsy which is invasive, destructive, and time-consuming. In this work, we propose and demonstrate the sample scanning nonlinear light microscopy, which is fast and non-invasive. By sample scanning microscope, the PVA concentrations between normal and abnormal films can be distinguished. By an axial scanning, the normal polarizer films have one absorption peak and the abnormal polarizer films have two absorption peaks. On the other hand, to boost up the imaging speed of a nonlinear light microscope, a beam scanning microscope was built up by Galvono mirrors. In the beam scanning nonlinear light microscopy, we use the Resolution Chart to definite resolution and image size. The best resolution was 0.87μm with a 250μm*250μm image size. Finally, we measure the two photon fluorescence images of mesophyll cells and chloroplast with different conditions.