TY - JOUR
T1 - A set of monomeric near-infrared fluorescent proteins for multicolor imaging across scales
AU - Matlashov, Mikhail E.
AU - Shcherbakova, Daria M.
AU - Alvelid, Jonatan
AU - Baloban, Mikhail
AU - Pennacchietti, Francesca
AU - Shemetov, Anton A.
AU - Testa, Ilaria
AU - Verkhusha, Vladislav V.
N1 - Funding Information:
We thank Olena Oliinyk (University of Helsinki, Finland) for the discussions and Michael Brenowitz (Albert Einstein College of Medicine, USA) for the analytical ultracentrifugation. This work was supported by the grants GM122567, NS103573 (both to V. V.V.) and EY030705 (to D.M.S.) from the US National Institutes of Health, and ERC-StG-638314 and 2016–03572 from the EU and Swedish research council (all to I.T.).
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - 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.
AB - 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.
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U2 - 10.1038/s41467-019-13897-6
DO - 10.1038/s41467-019-13897-6
M3 - Article
C2 - 31932632
AN - SCOPUS:85077785908
SN - 2041-1723
VL - 11
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 239
ER -