基于激光共聚焦显微镜λ扫描检测蒿草切片荧光光谱的研究

摘要

Objective To study the fluorescence spectrum of wormwood slice with laser confocal microscope,and provide the theoretical and experimental basis for obtaining the fluorescence spectrum of biomedical samples. Methods The fluorescence mi-croscope and laser scanning confocal microscope were used to observe fluorescence of wormwood slice. The experiment included three groups:group A,using fluorescence microscope to take photos;group B,using laser scanning confocal microscope to detect fluorescence spectrum;group C,using laser scanning confocal microscope to obtain fluorescence image. Results The fluorescence microscope could observe the fluorescence image of wormwood slice,but could not accurately obtain the corresponding fluores-cence excitation and emission wavelength. The channeling color interference phenomenon was occurred. The laser scanning confo-cal microscope could scan the fluorescence spectrum of wormwood slice,exclude the interference wavelength,obtain the fluores-cence excitation and emission wavelength accurately and gather fluorescent image with high-resolution. Conclusion Compared with the fluorescence microscope,the laser scanning confocal microscope can accurately obtain the fluorescence spectrum and high-resolution fluorescence image of biomedical samples.%目的：研究激光扫描共聚焦显微镜λ扫描检测蒿草切片的荧光光谱，为基于共聚焦显微镜获得生物医学样本的荧光光谱提供理论和实验依据。方法运用普通荧光显微镜和激光扫描共聚焦显微镜分别观察荧光染色的蒿草切片，实验分为三组：A组用普通荧光显微镜观察并拍照；B组用激光扫描共聚焦显微镜检测荧光光谱；C组用激光扫描共聚焦显微镜扫描荧光图像。结果荧光显微镜可以观察到蒿草切片的荧光图像，但不能准确获取其相应荧光激发和发射波长，而且会发生串色干扰现象。激光扫描共聚焦显微镜能对蒿草切片进行荧光光谱扫描，排除了干扰波长，获取其荧光激发和发射波长，得到高清晰度的荧光图像。结论与普通荧光显微镜比较，激光扫描共聚焦显微镜有能准确获取样本荧光光谱，得到高清晰度荧光图像的优点。