光谱法与共焦荧光成像法研究罗丹明B与ct-DNA的相互作用  

Study on Interaction between Rhodamine B and Calf Thymus DNA by Spectroscopy and Confocal Imaging

作　　者：于博昊[1] 缪文彬[2] 吴婷 康燕 姚建磊 张磊[1] 

机构地区：[1]华东理工大学化学与分子工程学院,上海200237 [2]上海出入境检验检疫局,上海200135

出　　处：《分析测试学报》2017年第9期1087-1092,共6页Journal of Instrumental Analysis

基　　金：国家质检总局科技计划项目(2016IK227)

摘　　要：采用吸收光谱、荧光光谱以及共焦荧光成像技术研究生理条件下罗丹明B(Rhodamine B,RB)与小牛胸腺DNA(ct-DNA)的相互作用以及相互作用模式。光谱研究结果显示,在450~650 nm的吸收光谱范围内,ct-DNA溶液对罗丹明B有减色作用;在560~660 nm荧光发射光谱范围内,ct-DNA对罗丹明B有荧光猝灭作用。同时,随着ct-DNA的加入,罗丹明B溶液的荧光偏振值发生变化,说明罗丹明B与ct-DNA能发生相互作用。在竞争性实验中,罗丹明B未能将亚甲基蓝(MB)从MB-ct-DNA复合体系中置换出来,说明罗丹明B与ct-DNA通过沟槽结合方式发生相互作用。共焦荧光成像结果显示,ct-DNA加入后,罗丹明B的荧光猝灭十分明显。利用罗丹明B和4',6-二脒基-2-苯基吲哚(DAPI)对He La细胞染色后进行共焦荧光成像,结果进一步证实罗丹明B与ct-DNA是通过沟槽结合作用将细胞核染成红色。The interaction between Rhodamine B and calf thymus DNA（ct-DNA） at physiological conditions were studied by the techniques of absorption spectroscopy, fluorescence spectroscopy and confocal imaging. The results of spectroscopy experiments suggested that the hypochromicity of Rhodamine B could appear in the presence of ct-DNA from 450nm to 650nm in the absorption spectroscopy, and the fluorescence of Rhodamine B was quenched in the presence of ct-DNA from 560nm to 660nm in the fluorescence emission spectroscopy. At the same time,the fluorescence polarization of Rhodamine B varied with the addition of ct-DNA.It was indicated that Rhodamine B could interact with ct-DNA. In the competition experiments, Rhodamine B was unable to replace methylene blue（MB） from MB-ct-DNA complex. It was indicated that the interaction between Rhodamine B and ct-DNA took place by the mode of groove combination. The results of confocal imaging experiments showed that the fluorescence of Rhodamine B was quenched quite obviously after the addition of ct-DNA. HeLa cells were dyed with Rhodamine B and 4′,6-diamidino-2-phenylindole（DAPI）,then used for confocal imaging. It was further verified that the cell nucleus could be dyed red through the groove combination interaction between Rhodamine B and ct-DNA.

关 键 词：罗丹明B(RB) 小牛胸腺DNA(ct-DNA) 沟槽结合 光谱法 共焦荧光成像 

分 类 号：O433.4[机械工程—光学工程] O641.3[理学—光学]