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Monomethine cyanine probes for visualization of cellular RNA by fluorescence microscopy
Methods and Applications in Fluorescence ( IF 3.2 ) Pub Date : 2021-07-13 , DOI: 10.1088/2050-6120/ac10ad
Daria Aristova 1, 2 , Viktoriia Kosach 1 , Svitlana Chernii 1, 2 , Yuriy Slominsky 3 , Anatoliy Balanda 1, 2 , Valeriy Filonenko 1 , Sergiy Yarmoluk 1, 2 , Alexandru Rotaru 4 , Hülya Gizem Özkan 5 , Andriy Mokhir 5 , Vladyslava Kovalska 1, 2
Affiliation  

We have studied spectral-luminescent properties of the monomethine cyanine dyes both in their free states and in the presence of either double-stranded deoxyribonucleic acids (dsDNAs) or single-stranded ribonucleic acids (RNAs). The dyes possess low fluorescence intensity in an unbound state, which is increased up to 479 times in the presence of the nucleic acids. In the presence of RNAs, the fluorescence intensity increase was stronger than that observed in the presence of dsDNA. Next, we have performed staining of live and fixed cells by all prepared dyes. The dyes proved to be cell and nuclear membrane permeant. They are photostable and brightly stain RNA-containing organelles in both live and fixed cells. The colocalization confirmed the specific nucleoli staining with anti-Ki-67 antibodies. The RNA digestion experiment has confirmed the selectivity of the dyes toward intracellular RNA. Based on the obtained results, we can conclude that the investigated monomethine cyanine dyes are useful fluorescent probes for the visualization of intracellular RNA and RNA-containing organelles such as nucleoli by using fluorescence microscopy.



中文翻译:

用于通过荧光显微镜观察细胞 RNA 的 Monomethine 花青探针

我们研究了单次甲基花青染料在游离状态和双链脱氧核糖核酸 (dsDNA) 或单链核糖核酸 (RNA) 存在下的光谱发光特性。染料在未结合状态下具有低荧光强度,在核酸存在时荧光强度增加了 479 倍。在 RNA 存在下,荧光强度增加比在 dsDNA 存在下观察到的更强。接下来,我们用所有制备的染料对活细胞和固定细胞进行了染色。染料被证明是细胞和核膜渗透性的。它们对活细胞和固定细胞中的含有 RNA 的细胞器具有光稳定性和明亮的染色效果。共定位证实了抗 Ki-67 抗体的特异性核仁染色。RNA 消化实验证实了染料对细胞内 RNA 的选择性。根据获得的结果,我们可以得出结论,所研究的单甲川花青染料是有用的荧光探针,可用于通过荧光显微镜观察细胞内 RNA 和含 RNA 的细胞器(如核仁)。

更新日期:2021-07-13
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