植物與人類的生活息息相關，不僅能夠乾淨的產生各種人類賴以生存的資源，對調節自然氣候或是水資源的保育都扮演著不可或缺的角色，故人類對植物學的知識建立是相當重要的。而在眾多的研究中，葉綠素螢光反應了相當程度的植物生理現象，但在早期對葉綠素螢光的研究上，往往需要透過化學層析的方式將葉綠素由搗碎的葉片內萃取出來，不僅嚴重的破壞了葉片內部的微結構，也降低了實驗環境與真實情況的符合程度。 隨著時代的演進，開始有科學家將光學顯微術引入了植物學的相關研究當中。本研究所採用的雙光子螢光顯微術，即是一套具優異光學切片能力、低破壞性之光學顯微技術，不需要對葉片進行樣本的處理即可進行葉綠素螢光的研究。以雙光子螢光顯微術的優勢為基礎，我們更進一步在影像系統架構中結合了光譜擷取系統，能在單次量測下分別記錄樣品的顯微影像及光譜訊號，解決了以往在光譜分析中，由於無法同時觀測到影像，故無法定確定螢光所發出位置的問題，並透過對十九種不同植物的影像及光譜分析，驗證了此一系統架構能作為植物研究上一套有利的量測及分析工具。; Plants are closely related to humans' daily life. It not only creates various living resources in a clean way but also plays an important role in weather regulation and water preservation. Therefore, it is necessary for human to establish the knowledge of botany. In many researches, chlorophyll fluorescence was found to reflect some significant plant physiological phenomenon. However, in the early researches of chlorophyll fluorescence, chlorophyll should be extracted from mashed leaves by chemical chromatography. These procedures not only destroy the micro-structures of leaves but also change the natural environment of leaves. With the evolution of techniques, optical microscopy began to be introduced in botany studies. In this research, two-photon fluorescence microscopy, which has optical sectioning power and low photodamages, is used to study the chlorophyll fluorescence without any sample preparation. Based on the advantages of two-photon fluorescence microscopy, a spectral detection system is further integrated with the imaging system to obtain the two-photon fluorescence image and the fluorescence spectrum in a single measurement. With the image information, it enables to make sure where the fluorescence comes from and realize the fluorescence distribution in the leaves. Through the analysis of images and spectra obtained from 19 different plants, this system is verified to be a useful tool for measurement and analysis in botany researches.
