Blue-shift photoconversion of near-infrared fluorescent proteins for labeling and tracking in living cells and organisms
Abstract Photolabeling of intracellular molecules is an invaluable approach to studying various dynamic processes in living cells with high spatiotemporal precision. Among fluorescent proteins, photoconvertible mechanisms and their products are in the visible spectrum (400–650 nm), limiting their in...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-12-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-44054-9 |
| _version_ | 1797376897907163136 |
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| author | Francesca Pennacchietti Jonatan Alvelid Rodrigo A. Morales Martina Damenti Dirk Ollech Olena S. Oliinyk Daria M. Shcherbakova Eduardo J. Villablanca Vladislav V. Verkhusha Ilaria Testa |
| author_facet | Francesca Pennacchietti Jonatan Alvelid Rodrigo A. Morales Martina Damenti Dirk Ollech Olena S. Oliinyk Daria M. Shcherbakova Eduardo J. Villablanca Vladislav V. Verkhusha Ilaria Testa |
| author_sort | Francesca Pennacchietti |
| collection | DOAJ |
| description | Abstract Photolabeling of intracellular molecules is an invaluable approach to studying various dynamic processes in living cells with high spatiotemporal precision. Among fluorescent proteins, photoconvertible mechanisms and their products are in the visible spectrum (400–650 nm), limiting their in vivo and multiplexed applications. Here we report the phenomenon of near-infrared to far-red photoconversion in the miRFP family of near infrared fluorescent proteins engineered from bacterial phytochromes. This photoconversion is induced by near-infrared light through a non-linear process, further allowing optical sectioning. Photoconverted miRFP species emit fluorescence at 650 nm enabling photolabeling entirely performed in the near-infrared range. We use miRFPs as photoconvertible fluorescent probes to track organelles in live cells and in vivo, both with conventional and super-resolution microscopy. The spectral properties of miRFPs complement those of GFP-like photoconvertible proteins, allowing strategies for photoconversion and spectral multiplexed applications. |
| first_indexed | 2024-03-08T19:45:13Z |
| format | Article |
| id | doaj.art-2d8a4d76185d4cd5bccf6a9421f0d8a7 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-08T19:45:13Z |
| publishDate | 2023-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-2d8a4d76185d4cd5bccf6a9421f0d8a72023-12-24T12:23:15ZengNature PortfolioNature Communications2041-17232023-12-0114111410.1038/s41467-023-44054-9Blue-shift photoconversion of near-infrared fluorescent proteins for labeling and tracking in living cells and organismsFrancesca Pennacchietti0Jonatan Alvelid1Rodrigo A. Morales2Martina Damenti3Dirk Ollech4Olena S. Oliinyk5Daria M. Shcherbakova6Eduardo J. Villablanca7Vladislav V. Verkhusha8Ilaria Testa9Department of Applied Physics and SciLifeLab, KTH Royal Institute of TechnologyDepartment of Applied Physics and SciLifeLab, KTH Royal Institute of TechnologyDivision of Immunology and Allergy, Department of Medicine Solna, Karolinska Institutet and University HospitalDepartment of Applied Physics and SciLifeLab, KTH Royal Institute of TechnologyDepartment of Applied Physics and SciLifeLab, KTH Royal Institute of TechnologyMedicum, University of HelsinkiDepartment of Genetics, and Gruss-Lipper Biophotonics Center, Albert Einstein College of MedicineDivision of Immunology and Allergy, Department of Medicine Solna, Karolinska Institutet and University HospitalMedicum, University of HelsinkiDepartment of Applied Physics and SciLifeLab, KTH Royal Institute of TechnologyAbstract Photolabeling of intracellular molecules is an invaluable approach to studying various dynamic processes in living cells with high spatiotemporal precision. Among fluorescent proteins, photoconvertible mechanisms and their products are in the visible spectrum (400–650 nm), limiting their in vivo and multiplexed applications. Here we report the phenomenon of near-infrared to far-red photoconversion in the miRFP family of near infrared fluorescent proteins engineered from bacterial phytochromes. This photoconversion is induced by near-infrared light through a non-linear process, further allowing optical sectioning. Photoconverted miRFP species emit fluorescence at 650 nm enabling photolabeling entirely performed in the near-infrared range. We use miRFPs as photoconvertible fluorescent probes to track organelles in live cells and in vivo, both with conventional and super-resolution microscopy. The spectral properties of miRFPs complement those of GFP-like photoconvertible proteins, allowing strategies for photoconversion and spectral multiplexed applications.https://doi.org/10.1038/s41467-023-44054-9 |
| spellingShingle | Francesca Pennacchietti Jonatan Alvelid Rodrigo A. Morales Martina Damenti Dirk Ollech Olena S. Oliinyk Daria M. Shcherbakova Eduardo J. Villablanca Vladislav V. Verkhusha Ilaria Testa Blue-shift photoconversion of near-infrared fluorescent proteins for labeling and tracking in living cells and organisms Nature Communications |
| title | Blue-shift photoconversion of near-infrared fluorescent proteins for labeling and tracking in living cells and organisms |
| title_full | Blue-shift photoconversion of near-infrared fluorescent proteins for labeling and tracking in living cells and organisms |
| title_fullStr | Blue-shift photoconversion of near-infrared fluorescent proteins for labeling and tracking in living cells and organisms |
| title_full_unstemmed | Blue-shift photoconversion of near-infrared fluorescent proteins for labeling and tracking in living cells and organisms |
| title_short | Blue-shift photoconversion of near-infrared fluorescent proteins for labeling and tracking in living cells and organisms |
| title_sort | blue shift photoconversion of near infrared fluorescent proteins for labeling and tracking in living cells and organisms |
| url | https://doi.org/10.1038/s41467-023-44054-9 |
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