Near-infrared bioluminescent proteins for two-color multimodal imaging

Bioluminescence imaging became a widely used technique for noninvasive study of biological processes in small animals. Bioluminescent probes with emission in near-infrared (NIR) spectral region confer the advantage of having deep tissue penetration capacity. However, there are a very limited number...

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Bibliographic Details
Published in:Scientific reports 2016-11, Vol.6 (1), p.36588-36588, Article 36588
Main Authors: Rumyantsev, Konstantin A., Turoverov, Konstantin K., Verkhusha, Vladislav V.
Format: Article
Language:English
Subjects:
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14/35
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631/1647/245/2222
631/1647/338/469
Bioluminescence
Chimeras
Color
Emissions
Energy transfer
Fluorescent indicators
Humanities and Social Sciences
I.R. radiation
Localization
Metastases
multidisciplinary
Probes
Science
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Summary:Bioluminescence imaging became a widely used technique for noninvasive study of biological processes in small animals. Bioluminescent probes with emission in near-infrared (NIR) spectral region confer the advantage of having deep tissue penetration capacity. However, there are a very limited number of currently available luciferases that exhibit NIR bioluminescence. Here, we engineered two novel chimeric probes based on RLuc8 luciferase fused with iRFP670 and iRFP720 NIR fluorescent proteins. Due to an intramolecular bioluminescence resonance energy transfer (BRET) between RLuc8 and iRFPs, the chimeric luciferases exhibit NIR bioluminescence with maxima at 670 nm and 720 nm, respectively. The 50 nm spectral shift between emissions of the two iRFP chimeras enables combined multicolor bioluminescence imaging (BLI) and the respective multicolor fluorescence imaging (FLI) of the iRFPs. We show that for subcutaneously implanted cells, NIR bioluminescence provided a 10-fold increase in sensitivity compared to NIR FLI. In deep tissues, NIR BLI enabled detection of as low as 10 4 cells. Both BLI and FLI allowed monitoring of tumor growth and metastasis from early to late stages. Multimodal imaging, which combines concurrent BLI and FLI, provides continuous spatiotemporal analysis of metastatic cells in animals, including their localization and quantification.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep36588