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Near-infrared fluorescent proteins for multicolor in vivo imaging

Nature Methods volume 10, pages 751–754 (2013)Cite this article

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Abstract

Near-infrared fluorescent proteins (FPs) are in high demand for in vivo imaging. We developed four spectrally distinct near-infrared FPs—iRFP670, iRFP682, iRFP702 and iRFP720—from bacterial phytochromes. iRFPs exhibit high brightness in mammalian cells and tissues and are suitable for long-term studies. iRFP670 and iRFP720 enable two-color imaging with standard approaches in living cells and mice. The four new iRFPs and the previously engineered iRFP713 allow multicolor imaging with spectral unmixing in living mice.

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Figure 1: Characterization of NIR FPs.
Figure 2: Multicolor imaging in vivo and in cells.

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Acknowledgements

We thank E. Giraud (Institute for Research and Development, France) for the hmuO and RpBphP6 genes, F. Subach (currently at the National Research Center “Kurchatov Institute”) for the pWA23h plasmid, G. Filonov (currently at Weill Cornell Medical College) for the iRFP713 adenovirus, J. Zhang for assistance in cell sorting, Y. Wang, L.-M. Ting, X. Wang and K. Piatkevich for help with mouse experiments, B. Taylor (PerkinElmer/Caliper) for assistance with the Living Image software and P. Guo for help with the confocal microscope. This work was supported by grants GM073913, CA164468 and EB013571 from the US National Institutes of Health.

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Authors and Affiliations

  1. Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York, USA

    Daria M Shcherbakova & Vladislav V Verkhusha

  2. Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, New York, USA

    Daria M Shcherbakova & Vladislav V Verkhusha

Authors
  1. Daria M Shcherbakova
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  2. Vladislav V Verkhusha
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Contributions

D.M.S. developed the proteins and characterized them in vitro, in cultured cells and in mice. V.V.V. directed and planned the project. V.V.V. and D.M.S. designed the experiments, analyzed the data and wrote the manuscript.

Corresponding author

Correspondence to Vladislav V Verkhusha.

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The authors declare no competing financial interests.

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Shcherbakova, D., Verkhusha, V. Near-infrared fluorescent proteins for multicolor in vivo imaging. Nat Methods 10, 751–754 (2013). https://doi.org/10.1038/nmeth.2521

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