一种数字化高灵敏度荧光显微镜及其在竹红菌甲素研究中的应用  

A Digitalized Ultra Sensitive Fluorescence Microscope and It′s Application in Hypocrellin

作　　者：陈天明[1] 王苏生[1] 俞信[1] 曹恩华[2,3] 许娜飞 李景福[2,3] 

机构地区：[1]北京理工大学工程光学系 [2]中国科学院生物物理研究所 [3]中国科学院上海技术物理研究所

出　　处：《光学学报》1996年第8期1157-1162,共6页Acta Optica Sinica

基　　金：国家自然科学基金

摘　　要：研制了一种可探测细胞微弱荧光图像的数字化高灵敏度荧光显微镜，采用视频数字化增强型ＣＣＤ系统作为高灵敏度的接收系统，像增强器亮度增益为４１，０００，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像，并可减少荧光物质的浓度和激发光的强度，减少对细胞自然生理环境的影响。数字化高灵敏度荧光显微镜在给出细胞微弱荧光图像的同时，并可给出图像上每一像元的发光强度和细胞平均发光强度。使用此仪器已首次直接得到了：１）光敏竹红菌甲素（ＨＡ）在ＨｅＬａ细胞（人体宫颈癌细胞）中的分布，证实ＨＡ主要分布在ＨｅＬａ的细胞膜中；２）ＨｅＬａ细胞在有ＨＡ存在时受强光照射后的光损伤现象；３）加入三种细胞抗氧化剂维生素Ｅ、丁羧基甲苯（ＳＦ）和阿魏酸钠（ＢＨＴ）后获得不同程度保护的细胞状态；４）一定浓度的ＨＡ在ＨｅＬａ细胞随时间的摄取曲线；５．不同ＨＡ浓度一定时间内在ＨｅＬａ细胞中被吸收量的曲线。A digitalized ultra sensitive fluorescence microscope has been developed, in which an intensifier CCD is used as high sensitive image detector with luminance gain of 41,000. It can detect ultra weak fluorescence image and decrease the concentration of fluorescence label and emitting light intensity in order to keep natural situation of the detected cell. This digitalized microscope can be used not only to obtain the fluorescence image, but also get the light luminance at every point of the image and the average light luminance of the whole image quantitatively. With this instrument, we have obtained: 1. The image of hypocrellin A distribution in HeLa cell, it shows that HA is mainly absorbed by cell membrane; 2. The image of HeLa cell be oxidated by strong light after adding HA; 3. Images of HeLa cell being protected by three antioxidant reagent Vit. E, sodium ferulate (SF) and butylated hydroxytoluene (BHT), from which it has been shown that anti oxidated effect of Vit E is the best; 4. The curve of absorbed quantity of HA in a HeLa cell with time versus culture time; 5. The curve of absorbed quantity of HA in Hela cell versus concentration of HA solution.

关 键 词：荧光显微镜 像增强器 光敏损伤 细胞抗氧化剂 

分 类 号：TH742.65[机械工程—光学工程] R912[机械工程—仪器科学与技术]