DNA staining for fluorescence and laser confocal microscopy
DNA staining for fluorescence and laser confocal microscopy
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DOI：
10.1177/002215549704500107
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发表时间：
1997-01-01
影响因子：
3.2
通讯作者：
Takata, K
中科院分区：
文献类型：
--
作者：
Suzuki, T;Fujikura, K;Takata, K
我们检查了五种核酸结合荧光染料，碘化丙啶、SYBR绿色I、YO-PRO-1、TOTO-3和TO-PRO-3，用于核DNA染色，通过荧光和激光共聚焦显微镜观察。研究了最佳浓度、RNA共染色和漂白速度。SYBR绿色I和TO-PRO-3几乎优先染色核DNA，而其他染料共染色胞质RNA。RNA酶处理完全防止了细胞质RNA染色。在传统的荧光显微镜中，这些染料可以与荧光标记的抗体组合使用。在测试的染料中，TOTO-3和TO-PRO-3在红色激发下用远红荧光染色DNA。在Kr/Ar激光照射下，TOTO-3和TO-PRO-3最适合作为荧光素和罗丹明（或德克萨斯红）免疫标记标本的核染色染料。
We examined five nucleic acid binding fluorescent dyes, propidium iodide, SYBR Green I, YO-PRO-1, TOTO-3, and TO-PRO-3, for nuclear DNA staining, visualized by fluorescence and laser confocal microscopy. The optimal concentration, co-staining of RNA, and bleaching speeds were examined. SYBR Green I and TO-PRO-3 almost preferentially stained the nuclear DNA, and the other dyes co-stained the cytoplasmic RNA. RNAse treatment completely prevented the cytoplasmic RNA staining. In conventional fluorescence microscopy, these dyes can be used in combination with fluorescence-labeled antibodies. Among the dyes tested, TOTO-3 and TO-PRO-3 stained the DNAs with far-red fluorescence under red excitation. Under Kr/Ar-laser illumination, TOTO-3 and TO-PRO-3 were best suited as the nuclear staining dyes in the specimens immunolabeled with fluorescein and rhodamine (or Texas red).