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A dendritic single-molecule fluorescent probe that is monovalent, photostable and minimally blinking

Nature Chemistry volume 5, pages 692–697 (2013)Cite this article

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Abstract

Single-molecule fluorescence techniques have emerged as a powerful approach to understanding complex biological systems. However, a challenge researchers still face is the limited photostability of nearly all organic fluorophores, including the cyanine and Alexa dyes. We report a new, monovalent probe that emits in the far-red region of the visible spectrum with properties desirable for single-molecule optical imaging. This probe is based on a ring-fused boron-dipyrromethene (BODIPY) core that is conjugated to a polyglycerol dendrimer (PGD). The dendrimer makes the hydrophobic fluorophore water-soluble. This probe exhibits excellent brightness, with an emission maximum of 705 nm. We have observed strikingly long and stable emission from individual PGD–BODIPY probes, even in the absence of anti-fading agents such as Trolox, a combined oxidizing–reducing agent often used in single-molecule studies for improving the photostability of common imaging probes. These interesting properties greatly simplify use of the fluorophore.

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Figure 1: Schematic representation of the synthesis of water-soluble and monofunctional polyglycerol-dendronized fluorophores.
Figure 2: Synthesis of biotinylated PGD–BODIPY 12.
Figure 3: MALDI-TOF mass spectra of 7–12.
Figure 4: Photophysical properties of 12.
Figure 5: Photostability comparison of 12 with Cy5.

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Acknowledgements

The authors acknowledge the National Institutes of Health (GM087448 to S.C.Z., GM065367 and AI083025 to T.H.) and the National Science Foundation (PHY-0822613) for financial support.

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Authors and Affiliations

  1. Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, 61801, Illinois, USA

    Si Kyung Yang & Steven C. Zimmerman

  2. Department of Physics, University of Illinois at Urbana-Champaign, Urbana, 61801, Illinois, USA

    Xinghua Shi, Seongjin Park & Taekjip Ha

  3. Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, 61801, Illinois, USA

    Xinghua Shi & Taekjip Ha

  4. Howard Hughes Medical Institute, University of Illinois at Urbana-Champaign, Urbana, 61801, Illinois, USA

    Xinghua Shi & Taekjip Ha

  5. Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, 61801, Illinois, USA

    Taekjip Ha

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  1. Si Kyung Yang
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Contributions

S.K.Y. and S.C.Z. conceived the study. S.K.Y. designed and synthesized the polyglycerol-dendronized fluorophores. X.S. performed the single-molecule experiments. X.S. and S.P. performed the microtubule labelling experiments. S.K.Y., X.S., T.H. and S.C.Z. wrote the manuscript.

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Correspondence to Taekjip Ha or Steven C. Zimmerman.

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Yang, S., Shi, X., Park, S. et al. A dendritic single-molecule fluorescent probe that is monovalent, photostable and minimally blinking. Nature Chem 5, 692–697 (2013). https://doi.org/10.1038/nchem.1706

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