A general strategy for developing cell-permeable photo-modulatable organic fluorescent probes for live-cell super-resolution imaging.
A general strategy for developing cell-permeable photo-modulatable organic fluorescent probes for live-cell super-resolution imaging.
复制标题
开发用于活细胞超分辨率成像的细胞渗透性光调制有机荧光探针的一般策略
DOI：
10.1038/ncomms6573
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发表时间：
2014-11-20
影响因子：
16.6
通讯作者：
Zhang, Yu-Hui
中科院分区：
文献类型：
--
作者：
Pan, Deng;Hu, Zhe;Qiu, Fengwu;Huang, Zhen-Li;Ma, Yilong;Wang, Yina;Qin, Lingsong;Zhang, Zhihong;Zeng, Shaoqun;Zhang, Yu-Hui
单分子定位显微镜（SMLM）实现了超越衍射极限的超分辨率成像，但严重依赖于光调制荧光探针的使用。在这里，我们报告了一个一般性的战略，通过利用显着的细胞溶质传递能力的细胞穿透肽（RR）3R 2构建细胞可渗透的光调制的有机荧光探针活细胞SMLM。我们开发了一种可光调制的有机荧光探针，它由一个（RR）3R 2肽与一个细胞不可渗透的有机荧光团和一个识别单元组成。我们的研究结果表明，这些有机探针不仅是细胞渗透，但也可以特异性和直接标记内源性靶蛋白。使用探针，我们获得超分辨率图像的溶酶体和内源性F-肌动蛋白在生理条件下。我们在活细胞中以10 s的时间分辨率解决了F-肌动蛋白的动力学，并分辨出直径约80 nm的精细F-肌动蛋白结构。这些结果为活细胞超分辨率成像的荧光探针的设计开辟了新的途径。
Single-molecule localization microscopy (SMLM) achieves super-resolution imaging beyond the diffraction limit but critically relies on the use of photo-modulatable fluorescent probes. Here we report a general strategy for constructing cell-permeable photo-modulatable organic fluorescent probes for live-cell SMLM by exploiting the remarkable cytosolic delivery ability of a cell-penetrating peptide (rR)3R2. We develop photo-modulatable organic fluorescent probes consisting of a (rR)3R2peptide coupled to a cell-impermeable organic fluorophore and a recognition unit. Our results indicate that these organic probes are not only cell permeable but can also specifically and directly label endogenous targeted proteins. Using the probes, we obtain super-resolution images of lysosomes and endogenous F-actin under physiological conditions. We resolve the dynamics of F-actin with 10 s temporal resolution in live cells and discern fine F-actin structures with diameters of ~80 nm. These results open up new avenues in the design of fluorescent probes for live-cell super-resolution imaging.
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影响因子：
15
作者：
Lee, Hsiao-lu D.;Lord, Samuel J.;Iwanaga, Shigeki;Zhan, Ke;Xie, Hexin;Williams, Jarrod C.;Wang, Hui;Bowman, Grant R.;Goley, Erin D.;Shapiro, Lucy;Twieg, Robert J.;Rao, Jianghong;Moerner, W. E.
通讯作者：
Moerner, W. E.
影响因子：
48
作者：
Dempsey, Graham T.;Vaughan, Joshua C.;Chen, Kok Hao;Bates, Mark;Zhuang, Xiaowei
通讯作者：
Zhuang, Xiaowei
影响因子：
4.7
作者：
Mueller, Judith;Kretzschmar, Ines;Boisguerin, Prisca
通讯作者：
Boisguerin, Prisca
影响因子：
4.7
作者：
Fan, Fengkai;Nie, Si;Zhang, Yu-Hui
通讯作者：
Zhang, Yu-Hui
影响因子：
48
作者：
Riedl, Julia;Crevenna, Alvaro H.;Wedlich-Soldner, Roland
通讯作者：
Wedlich-Soldner, Roland