On-Target Photoassembly of Pyronin Dyes for Super-Resolution Microscopy.

IF 16.9
Gergely Knorr, Mariano L Bossi, Stefan W Hell
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Abstract

Controlled photoactivation is an auspicious and emerging approach in super-resolution microscopy, offering virtually zero background signal from the marker prior to activation. Pyronins are well-established fluorophores, but due to their inherent intercalating tendency towards nucleic acids, their use has been mostly avoided in super-resolution microscopy. Here, we describe a new class of diaryl ether and diaryl silane molecules that upon photoactivation close into fluorescent (silicon-)pyronins and term them Pyronin Upon Light Irradiation (PULI). This concept exploits the outstanding photophysical properties of pyronins (bright, photostable, and optimal spectral features for standard microscopes), while overcoming their major drawback (intrinsic affinity of accumulating in the nucleus and around RNA) for the design of fluorescent markers for imaging applications. Furthermore, we also demonstrate that this approach is applicable to their Si-bridged analogues, extending this family of photoactivatable molecules to the far-red regime. The versatility of our approach was also highlighted by tagging diverse biological targets in cells and visualizing them using advanced super-resolution microscopy techniques, such as PALM, STED, and MINFLUX.

超分辨显微镜中Pyronin染料的靶光组装。
可控光激活是一种吉祥的和新兴的方法在超分辨率显微镜,提供几乎零背景信号从激活之前的标记。吡咯蛋白是公认的荧光团,但由于其固有的嵌入倾向于核酸,它们的使用大多避免在超分辨率显微镜。在这里,我们描述了一类新的二芳基醚和二芳基硅烷分子,它们在光激活后接近荧光(硅-)Pyronin,并将其命名为光照射Pyronin (PULI)。这个概念利用了pyronins杰出的光物理特性(明亮,光稳定,标准显微镜下的最佳光谱特征),同时克服了它们的主要缺点(在细胞核和RNA周围积累的内在亲和力),用于设计用于成像应用的荧光标记。此外,我们还证明了这种方法适用于他们的硅桥类似物,将这一家族的光激活分子扩展到远红色区域。通过标记细胞中的各种生物靶标并使用先进的超分辨率显微镜技术(如PALM, STED和MINFLUX)将其可视化,我们的方法的多功能性也得到了强调。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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