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

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 l...

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Bibliographic Details
Published in:Nature communications 2014-11, Vol.5 (1), p.5573-5573, Article 5573
Main Authors: Pan, Deng, Hu, Zhe, Qiu, Fengwu, Huang, Zhen-Li, Ma, Yilong, Wang, Yina, Qin, Lingsong, Zhang, Zhihong, Zeng, Shaoqun, Zhang, Yu-Hui
Format: Article
Language:English
Subjects:
631/1647/1888
631/80/2373/2238
Actins - ultrastructure
Cell-Penetrating Peptides - chemistry
Fluorescent Dyes - chemistry
Humanities and Social Sciences
Humans
Lysosomes - ultrastructure
Microscopy, Fluorescence
multidisciplinary
Nanotechnology
Science
Science (multidisciplinary)
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Summary: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 ~80 nm. These results open up new avenues in the design of fluorescent probes for live-cell super-resolution imaging. Single-molecule localization microscopy depends on the use of photo-modulatable fluorescent probes; however, many cannot be used in live-cell studies due to poor cell permeability. Pan et al. present a strategy for constructing cell-permeable probes and use it to image actin filament dynamics and lysosomes.
ISSN:2041-1723
2041-1723
DOI:10.1038/ncomms6573