Near-infrared bioluminescent proteins for two-color multimodal imaging.
Near-infrared bioluminescent proteins for two-color multimodal imaging.
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DOI：
10.1038/srep36588
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发表时间：
2016-11-11
影响因子：
4.6
通讯作者：
Verkhusha VV
中科院分区：
文献类型：
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作者：
Rumyantsev KA;Turoverov KK;Verkhusha VV
生物发光成像技术已成为一种广泛应用于小动物生物过程非侵入性研究的技术。具有近红外（NIR）光谱区域中的发射的生物发光探针赋予具有深组织穿透能力的优点。然而，目前可获得的表现出NIR生物发光的酶的数量非常有限。在这里，我们设计了两种新的嵌合探针，其基于与iRFP670和iRFP720 NIR荧光蛋白融合的RLuc8荧光素酶。由于RLuc8和iRFPs之间的分子内生物发光共振能量转移（BRET），嵌合体酶表现出分别在670 nm和720 nm处具有最大值的NIR生物发光。两种iRFP嵌合体的发射之间的50 nm光谱偏移使得能够实现iRFP的组合的生物发光成像（BLI）和各自的荧光成像（FLI）。我们发现，对于皮下植入的细胞，近红外生物发光提供了10倍的灵敏度增加相比，近红外FLI。在深层组织中，NIR BLI能够检测低至104个细胞。BLI和FLI都可以从早期到晚期监测肿瘤生长和转移。多模式成像结合了同步BLI和FLI，可对动物转移性细胞进行连续时空分析，包括其定位和定量。
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 104 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.
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影响因子：
11.2
作者：
Schaub FX;Reza MS;Flaveny CA;Li W;Musicant AM;Hoxha S;Guo M;Cleveland JL;Amelio AL
通讯作者：
Amelio AL
影响因子：
4.3
作者：
Mezzanotte, Laura;Blankevoort, Vicky;Lowik, Clemens W. G. M.;Kaijzel, Eric L.
通讯作者：
Kaijzel, Eric L.
影响因子：
--
作者：
Shcherbakova DM;Baloban M;Pletnev S;Malashkevich VN;Xiao H;Dauter Z;Verkhusha VV
通讯作者：
Verkhusha VV
影响因子：
7.1
作者：
Xia, J. J.;Berg, E. P.;Yao, G.
通讯作者：
Yao, G.
影响因子：
16.6
作者：
通讯作者：
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