本論文是以雙光子激發受激輻射耗損顯微鏡為目標,討論環形光場分布以及樣品螢光生命週期對其解析度之影響,提出增加解析度的方法。在環形光場的討論中,本研究採用模擬及光場量測分析以不同方式所產生的環形光場及其在強聚焦下的光場變化,而螢光生命週期的研究則是模擬的方式來了解螢光樣品的選擇可以如何幫助解析度的提昇。此外,本研究已著手進行系統的建構,期望能透過實際系統的建構來證實上述論點之可行性。 This thesis is based on the two-photon excitation stimulated-emission depletion microscopy to discuss the effects of donut beam and fluorescence lifetime on the spatial resolution of this system and propose the methods to improve the resolution. In this research, both simulation and experimental measurements were applied to analyze the intensity distributions of donut beams generated in different ways and their distributions under tightly-focusing condition. On the other hand, only simulation was used to discuss the impact of fluorescence lifetime on the spatial resolution improvement. Finally, in order to obtain the experimental evidences of our results, the two-photon excitation stimulated-emission depletion microscopy was setup.