高灵敏度荧光显微镜量化技术及其在光生物学研究中的应用

介绍一种用像增器接收荧光图像的高灵敏度荧光显微镜，相对于普通荧光显微镜的灵敏度提高了４×１０４倍，并用宽量程微光光亮度计对仪器的微弱成像性能进行了实验标定，得到了图像采集数据和图像发光强度的线性数量关系。高灵敏度荧光显微镜在给出细胞荧光图像的同时，可以给出图像上每一像元的发光强度和细胞平均发光强度，仪器对图像细微变化的判断能力远大于人眼直接观察图像。高灵敏度荧光显微镜可应用于研究细胞中荧光物质在细胞生理过程中的分布变化和发光强度变化。使用此仪器已得到了光敏竹红菌甲素（ＨＡ）在Ｈｅｌａ细胞（人体子宫癌细胞）中的分布图像和更为直观的三维显示图形，以及加入ＨＡ后Ｈｅｌａ细胞受到强先照射后的细胞损伤图像。

DIGITALIZATION IN ULTRA-SENSITIVE FLUOPESENCE MICROSCOPY AND ITS APPLICATION IN PHOTO-BIOLOGY RESEARCH

An ultra-sensitive fluorescence microsocopy has been developed with the image intensifier as its detector. Its sensitivity is 4 ×104 higher than that of ordinary fluorescence microscopy, and the linear coefficiency between light intensity and CCD output was obtained by using a wide-range ultra-weak photometer. So the instrument can not only show the image of ultra-weak fluoresrmce distribution, but also give the intensities of fluorescence of each iTnage panel as well as that of a Cell. Thus, the instrument can be used to detect trivial changing of light intenSity or light distribution which is toossible to be obserVed with human eye directly.Using this ulha-sensitive fluorescence microscopy, we have got the image of Hypocrellin A distribution in Hela Cells and its three dimensional display, we have also got the cage of Hela Cell being oxidated after exposed to strong light.