A general strategy for developing cell-permeable photo-modulatable organic fluorescent probes for live-cell super-resolution imaging

作者：Pan, Deng; Hu, Zhe; Qiu, Fengwu; Huang, Zhen-Li; Ma, Yilong; Wang, Yina; Qin, Lingsong; Zhang, Zhihong; Zeng, Shaoqun; Zhang, Yu-Hui^*

来源：Nature Communications, 2014, 5(1): 5573.

DOI：10.1038/ncomms6573

• PHOTOACTIVATION LOCALIZATION MICROSCOPY
• RESOLUTION LIMIT
• ACTIN
• FLUOROPHORES
• NANOSCALE
• PROTEINS

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)(3)R-2. We develop photo-modulatable organic fluorescent probes consisting of a (rR)(3)R-2 peptide 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 similar to 80 nm. These results open up new avenues in the design of fluorescent probes for live-cell super-resolution imaging.

• 出版日期2014-11
• 单位华中科技大学