A set of monomeric near-infrared fluorescent proteins for multicolor imaging across scales
| Autor: | Daria M. Shcherbakova, Anton A. Shemetov, Vladislav V. Verkhusha, Jonatan Alvelid, Francesca Pennacchietti, Ilaria Testa, Mikhail E. Matlashov, Mikhail Baloban |
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| Přispěvatelé: | Medicum, Faculty of Medicine, University of Helsinki |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Brightness Intravital Microscopy General Physics and Astronomy Protein tag Protein Engineering Fluorescence imaging Mice 0302 clinical medicine Microscopy Super-resolution microscopy lcsh:Science BRIGHT Spectroscopy Near-Infrared Multidisciplinary Protein Stability Biological techniques Resolution (electron density) STED microscopy MICROSCOPY Fluorescence Molecular Imaging INSIGHTS LIGHT surgical procedures operative Female Preclinical imaging Materials science animal structures Science Article General Biochemistry Genetics and Molecular Biology Cell Line 03 medical and health sciences Animals Humans neoplasms KNOT Near-infrared spectroscopy technology industry and agriculture General Chemistry equipment and supplies Luminescent Proteins 030104 developmental biology Biophysics lcsh:Q 3111 Biomedicine Protein design 030217 neurology & neurosurgery |
| Zdroj: | Nature Communications, Vol 11, Iss 1, Pp 1-12 (2020) Nature Communications |
| ISSN: | 2041-1723 |
| Popis: | 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. Monomeric near-infrared (NIR) fluorescent proteins (FPs) from bacterial phytochromes bring potential advantages, but their brightness in cells is lower than dimeric NIR FPs. Here the authors develop enhanced monomeric NIR FPs enabling imaging across different scales without the trade-off between brightness and monomeric state. |
| Databáze: | OpenAIRE |
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