Ultra-photostable DNA FluoroCubes: Mechanism of Photostability and Compatibility with FRET and Dark Quenching.
Ultra-photostable DNA FluoroCubes: Mechanism of Photostability and Compatibility with FRET and Dark Quenching.
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DOI：
10.1021/acs.nanolett.2c01757
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发表时间：
2022-08-10
期刊：
影响因子：
10.8
通讯作者：
Walter, Nils G.
中科院分区：
文献类型：
--
作者：
Blanchard, Aaron T.;Li, Zi;Duran, Elizabeth C.;Scull, Catherine E.;Hoff, J. Damon;Wright, Keenan R.;Pan, Victor;Walter, Nils G.
基于 DNA 的 FluoroCube 最近被开发为光漂白的解决方案，光漂白是荧光显微镜普遍存在的局限性（Niekamp、Stuurman 和 Vale，NatureMethods，2020）。 FluoroCubes – 由 ≤6 个荧光染料结合的紧凑型 ~4×4×5.4nm3 四螺旋束 – 保持荧光的时间比单一染料长约 50 倍，并发射高达约 40 倍的光子。这项工作回答了有关 FluoroCubes 的两个重要问题：首先，增强光稳定性的机制是什么？其次，FluoroCube 是否与福斯特共振能量转移 (FRET) 和类似技术兼容？在这里，我们使用单粒子光漂白研究来证明光稳定性是通过荧光团和四螺旋 DNA 束之间的相互作用产生的。支持这一点的是，我们发现较小的~4×4×2.7 nm3 FluoroCubes也具有超强的光稳定性。然而，我们发现某些染料与染料之间的相互作用会对 FluoroCube 的性能产生负面影响。因此，缺乏这些相互作用的 4 染料 FluoroCube 比 6 染料 FluoroCube 表现更好。我们还证明 FluoroCube 与 FRET 和暗淬火应用兼容。
DNA-based FluoroCubes were recently developed as a solution to photobleaching, a ubiquitous limitation of fluorescence microscopy (Niekamp, Stuurman, & Vale, Nature Methods, 2020). FluoroCubes – compact ~4×4×5.4nm3 four-helix bundles bound by ≤6 fluorescent dyes – remain fluorescent up to ~50× longer than single dyes and emit up to ~40× as many photons. This work answers two important questions about FluoroCubes: First, what is the mechanism by which photostability is enhanced? Second, are FluoroCubes compatible with Förster resonance energy transfer (FRET) and similar techniques? Here, we use single particle photobleaching studies to show that photostability arises through interactions between the fluorophores and the four-helix DNA bundle. Supporting this, we discover that smaller ~4×4×2.7 nm3 FluoroCubes also confer ultra-photostability. However, we find that certain dye-dye interactions negatively impact FluoroCube performance. Accordingly, 4-dye FluoroCubes lacking these interactions perform better than 6-dye FluoroCubes. We also demonstrate that FluoroCubes are compatible with FRET and dark quenching applications.
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影响因子：
48
作者：
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通讯作者：
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影响因子：
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通讯作者：
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作者：
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通讯作者：
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