高灵敏度荧光显微镜及其在细胞光敏效应研究中的应用

Ultra-Sensitive Fluorescence Microscope and Its Applications in Cell Photosensitive Effect

研制了一种可探测极微弱细胞荧光图象的高灵敏度荧光显微镜，在仪器中使用了一个高质量的亮度增益为５０，０００的象增强器对细胞的荧光图象进行光能量放大，因而这种荧光显微镜可探测到普通荧光显微镜不能观察的微弱荧光图象。由于仪器中光源的输出强度很小，避免了长时间观察对活细胞的光损伤，并可观察到荧光效率低的荧光物质的分布图象。探测系统中用计算机采集图象数据，同时对数据进行必要处理，以提高图象的质量。这种高灵敏度的荧光显微镜可用于分析细胞中荧光物质的分布，以及细胞动态生理过程的研究。使用此仪器已直接观察到光敏竹红菌甲素在Ｈｅｌａ细胞（人体宫颈癌细胞）中的分布，细胞受光照射后的光损伤现象及加入三种细胞保护剂后获得不同程度保护的细胞状态。

An ultra-sensitive fluorescence microscope has been developed. It can detect ultra-weak fluorescence image of cells in cell biology research. In this instrument, a high quality image intensifier with luminance gain of 50,000 is used as an image detector for imaging of the ultra-weak fluorescence of cells. This instrument is much more sensitive than common fluorescence imcroscope, it can obtain images which are impossible by using an ordinary fluorescence microscope due to low concentration of fluorescence label material or photo-bleaching. A personal computer and some image processing methods are used to improve the image quality and calculate quantitative parameters. This instrument can detect the distribution of fluorescence label materials in all kinds of cells, and it can also detect the information of dynamic process in cell biology research. With this instrument we have directly observed the distribution of Hypocrellin A in Hela cells , the cells damaging caused by strong light emitting and different protective conditions of cells after adding three different cell protective reagents.