Latt S A, Marino M, Lalande M
Cytometry. 1984 Jul;5(4):339-47. doi: 10.1002/cyto.990050409.
The spectroscopic properties of three new dyes, EK4, VL772, and LL585, free and bound to nucleic acids, are presented, with particular emphasis on their potential use in flow cytometry. Two of these dyes, EK4 and LL585, exhibit red fluorescence, while dye VL772 exhibits yellow fluorescence. Dye LL585 exhibits specificity for DNA, relative to RNA, and a marked enhancement of fluorescence efficiency upon binding to DNA, needed for a red fluorescent DNA-specific stain for flow cytometry. The dye penetrates live cells, although uniformity of nuclear fluorescence, as evidenced by DNA flow histograms, is better if the cells are first permeabilized with Triton X-100. Dye VL772 exhibits yellow fluorescence and little DNA-RNA discrimination, but may prove useful in conjunction with dye LL585 when simultaneous assay of cellular RNA and DNA is desired. Dye EK4 shares properties of the other two dyes but fluoresces with much less efficiency. Dyes LL585 and VL772, used singly, as a pair, or in combination with blue-fluorescing DNA specific dyes, such as bisbenzimidazole derivatives, should permit new, convenient analyses of the content and organization of cellular nucleic acids.
本文介绍了三种新染料EK4、VL772和LL585游离状态及与核酸结合状态下的光谱特性，特别强调了它们在流式细胞术中的潜在应用。其中两种染料EK4和LL585发出红色荧光，而染料VL772发出黄色荧光。相对于RNA，染料LL585对DNA具有特异性，并且在与DNA结合时荧光效率显著增强，这是流式细胞术红色荧光DNA特异性染色所必需的。该染料可穿透活细胞，不过，若先用Triton X-100使细胞通透化，如DNA流式直方图所示，核荧光的均匀性会更好。染料VL772发出黄色荧光，对DNA和RNA的区分能力较弱，但在需要同时检测细胞RNA和DNA时，与染料LL585联合使用可能会很有用。染料EK4具有其他两种染料的特性，但荧光效率低得多。染料LL585和VL772单独使用、成对使用或与蓝色荧光DNA特异性染料（如双苯并咪唑衍生物）联合使用，应该能够对细胞核酸的含量和组织进行新的、便捷的分析。
