RESOLFT nanoscopy with photoswitchable organic fluorophores.
RESOLFT nanoscopy with photoswitchable organic fluorophores.
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DOI：
10.1038/srep17804
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发表时间：
2015-12-07
影响因子：
4.6
通讯作者：
Kim SK
中科院分区：
文献类型：
--
作者：
Kwon J;Hwang J;Park J;Han GR;Han KY;Kim SK
远场光学纳米镜已广泛用于以亚衍射极限空间分辨率对小物体进行成像。特别是，使用光可切换荧光蛋白的可逆饱和光学荧光跃迁（RESOLFT）纳米镜是一种在低光强度下对活细胞进行超分辨率成像的强大方法。在这里，我们首次展示了使用有机荧光团在生物系统中实施 RESOLFT 纳米显微镜，有机荧光团尺寸更小，更容易进行化学修饰。通过共价连接的 Cy3 和 Alexa647 染料对来标记固定细胞中的亚细胞结构，并通过优化成像缓冲区和光学参数，我们的 RESOLFT 纳米镜在焦平面上实现了约 74 nm 的空间分辨率。该方法为低光强度 RESOLFT 纳米显微镜提供了强大的替代方案，可以使用小型有机探针以纳米级分辨率进行生物成像。
Far-field optical nanoscopy has been widely used to image small objects with sub-diffraction-limit spatial resolution. Particularly, reversible saturable optical fluorescence transition (RESOLFT) nanoscopy with photoswitchable fluorescent proteins is a powerful method for super-resolution imaging of living cells with low light intensity. Here we demonstrate for the first time the implementation of RESOLFT nanoscopy for a biological system using organic fluorophores, which are smaller in size and easier to be chemically modified. With a covalently-linked dye pair of Cy3 and Alexa647 to label subcellular structures in fixed cells and by optimizing the imaging buffer and optical parameters, our RESOLFT nanoscopy achieved a spatial resolution of ~74 nm in the focal plane. This method provides a powerful alternative for low light intensity RESOLFT nanoscopy, which enables biological imaging with small organic probes at nanoscale resolution.