共焦顯微鏡系統之建構與實驗量測

Abstract

為了在實驗中觀測微細物件，且鑒於直接購買高解析度共焦式顯微鏡昂貴及無法隨意拆組特性，在實驗室中建構了共焦式雷射掃描顯微鏡系統，其可用於掃描金屬樣品和螢光樣品。目前顯微鏡系統使用波長為473 nm的藍光雷射，以及NA 0.8之物鏡，並可觀測到直徑長100 nm的螢光乳膠粒。 本系統以LabVIEW™工具編寫系統控制程式以整合不同光學元件以及機械移動平台，並利用計算機結合其點資訊以輸出圖像。在取得影像後以MATLAB®工具建立影像重構程式嘗試由掃描影像回解出點擴散函數及原始物體形貌。

A laser scanning confocal microscope was developed in this study to examine microstructures without incurring the high cost of purchasing a microsope. The confocal microscope uses a 473 nm laser as the light source and employs a high numerical aperture objective (NA = 0.8).It can be used to examine metallic samples with a 200 nm resolution and fluorescence samples with a 100 nm resolution. The confocal microscope is operated with a program coded in LabVIEW™. The program integrates optical elements with the positioning stage, which is connected to the computer. In addition, a preliminary program that restores objects and has a point spread function was developed in this study by using MATLAB®.