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A set of monomeric near-infrared fluorescent proteins for multicolor imaging across scales

Author

Listed:
  • Mikhail E. Matlashov

    (Department of Anatomy and Structural Biology and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx)

  • Daria M. Shcherbakova

    (Department of Anatomy and Structural Biology and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx)

  • Jonatan Alvelid

    (Department of Applied Physics and Science for Life Laboratory, KTH Royal Institute of Technology)

  • Mikhail Baloban

    (Department of Anatomy and Structural Biology and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx)

  • Francesca Pennacchietti

    (Department of Applied Physics and Science for Life Laboratory, KTH Royal Institute of Technology)

  • Anton A. Shemetov

    (Department of Anatomy and Structural Biology and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx)

  • Ilaria Testa

    (Department of Applied Physics and Science for Life Laboratory, KTH Royal Institute of Technology)

  • Vladislav V. Verkhusha

    (Department of Anatomy and Structural Biology and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx
    Medicum, Faculty of Medicine, University of Helsinki)

Abstract

Bright monomeric near-infrared (NIR) fluorescent proteins (FPs) are in high demand as protein tags for multicolor microscopy and in vivo imaging. Here we apply rational design to engineer a complete set of monomeric NIR FPs, which are the brightest genetically encoded NIR probes. We demonstrate that the enhanced miRFP series of NIR FPs, which combine high effective brightness in mammalian cells and monomeric state, perform well in both nanometer-scale imaging with diffraction unlimited stimulated emission depletion (STED) microscopy and centimeter-scale imaging in mice. In STED we achieve ~40 nm resolution in live cells. In living mice we detect ~105 fluorescent cells in deep tissues. Using spectrally distinct monomeric NIR FP variants, we perform two-color live-cell STED microscopy and two-color imaging in vivo. Having emission peaks from 670 nm to 720 nm, the next generation of miRFPs should become versatile NIR probes for multiplexed imaging across spatial scales in different modalities.

Suggested Citation

  • Mikhail E. Matlashov & Daria M. Shcherbakova & Jonatan Alvelid & Mikhail Baloban & Francesca Pennacchietti & Anton A. Shemetov & Ilaria Testa & Vladislav V. Verkhusha, 2020. "A set of monomeric near-infrared fluorescent proteins for multicolor imaging across scales," Nature Communications, Nature, vol. 11(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-019-13897-6
    DOI: 10.1038/s41467-019-13897-6
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    Cited by:

    1. Yuang Gu & Jingqu Chen & Zhaoran Wang & Chang Liu & Tianzheng Wang & Chan-Jin Kim & Helena Durikova & Soraia Fernandes & Darryl N. Johnson & Robert De Rose & Christina Cortez-Jugo & Frank Caruso, 2024. "mRNA delivery enabled by metal–organic nanoparticles," Nature Communications, Nature, vol. 15(1), pages 1-15, December.

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