Suzuki T, Fujikura K, Higashiyama T, Takata K
Department of Cell Biology, Gunma University, Japan.
J Histochem Cytochem. 1997 Jan;45(1):49-53. doi: 10.1177/002215549704500107.
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).
我们检测了五种核酸结合荧光染料，碘化丙啶、SYBR Green I、YO-PRO-1、TOTO-3和TO-PRO-3，用于细胞核DNA染色，通过荧光和激光共聚焦显微镜进行观察。检测了最佳浓度、RNA共染色情况以及漂白速度。SYBR Green I和TO-PRO-3几乎优先对细胞核DNA进行染色，而其他染料则对细胞质RNA进行共染色。RNA酶处理完全阻止了细胞质RNA染色。在传统荧光显微镜下，这些染料可与荧光标记抗体联合使用。在所测试的染料中，TOTO-3和TO-PRO-3在红色激发下用远红色荧光对DNA进行染色。在Kr/Ar激光照射下，TOTO-3和TO-PRO-3最适合作为在用荧光素和罗丹明（或德克萨斯红）进行免疫标记的标本中的细胞核染色染料。
