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

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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 digital high-sensitivity fluorescence microscope was developed to detect the weak fluorescence images of cells. Using a digital video enhanced CCD system as a high-sensitivity receiving system with an image intensifier gain of 41,000, the fluorescence microscope can detect normal Fluorescence microscope can not observe the weak fluorescence images, and can reduce the concentration of fluorescent substances and the intensity of the excitation light to reduce the impact on the cell’s natural physiological environment. Digital high-sensitivity fluorescence microscopy gives the cells a weak fluorescence image at the same time, and gives the image of each pixel luminescence intensity and the average luminous intensity of the cell. For the first time, this instrument has been used directly: 1) The distribution of Hypocrellin A (HA) in HeLa cells (human cervical cancer cells), confirming that HA is mainly distributed in the cell membrane of HeLa; 2) 3) adding three kinds of cell antioxidants such as vitamin E, butylated hydroxytoluene (SF) and sodium ferulate (BHT) to obtain different degrees of protection of the cell state; 4) a certain concentration Of HA uptake in HeLa cells over time; 5. Absorbance in HeLa cells at different HA concentrations over time.
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