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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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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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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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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DOI: https://doi.org/10.1038/nchem.1706
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