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

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  • Deng Pan

    (Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology
    Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology)

  • Zhe Hu

    (Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology
    Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology)

  • Fengwu Qiu

    (Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology
    Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology)

  • Zhen-Li Huang

    (Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology
    Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology)

  • Yilong Ma

    (Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology
    Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology)

  • Yina Wang

    (Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology
    Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology)

  • Lingsong Qin

    (Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology
    Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology)

  • Zhihong Zhang

    (Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology
    Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology)

  • Shaoqun Zeng

    (Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology
    Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology)

  • Yu-Hui Zhang

    (Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology
    Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology)

Abstract

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)3R2 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 ~80 nm. These results open up new avenues in the design of fluorescent probes for live-cell super-resolution imaging.

Suggested Citation

  • Deng Pan & Zhe Hu & Fengwu Qiu & Zhen-Li Huang & Yilong Ma & Yina Wang & Lingsong Qin & Zhihong Zhang & Shaoqun Zeng & Yu-Hui Zhang, 2014. "A general strategy for developing cell-permeable photo-modulatable organic fluorescent probes for live-cell super-resolution imaging," Nature Communications, Nature, vol. 5(1), pages 1-8, December.
    • Handle: RePEc:nat:natcom:v:5:y:2014:i:1:d:10.1038_ncomms6573
    DOI: 10.1038/ncomms6573
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