Luminescent proteins for high-speed single-cell and whole-body imaging.
Luminescent proteins for high-speed single-cell and whole-body imaging.
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DOI：
10.1038/ncomms2248
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发表时间：
2012
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
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荧光蛋白的使用彻底改变了我们对生物过程的理解。然而，对外部照明的要求排除了它们对所有组织中生物过程的研究的普遍应用。虽然光可以通过化学发光产生，但是来自现有化学发光探针的光发射太弱而不能在不能使用荧光的情况下使用这种成像模式。在这里，我们报告了迄今为止最亮的发光蛋白的发展，纳米灯笼，这是增强海肾荧光素酶和金星，具有高生物发光共振能量转移效率的荧光蛋白的嵌合体。Nano-lantern允许活细胞中的细胞内结构的实时成像，其空间分辨率相当于荧光和自由移动的未剃毛小鼠中的灵敏肿瘤检测。我们还创建了基于Nano-lantern的功能指示剂，可以在无法使用荧光指示剂的环境中对Ca 2+、环磷酸腺苷和5′-三磷酸腺苷动态进行成像。这些发光蛋白质允许在动物和植物中以前看不见的单细胞，器官和全身水平上可视化生物现象。 发光蛋白是生物医学成像的重要工具，但往往发出相当少的光。Saito等人描述了一种更明亮的生物发光蛋白，它可以以高的空间和时间分辨率显示各种信号分子的细胞内动力学。
The use of fluorescent proteins has revolutionized our understanding of biological processes. However, the requirement for external illumination precludes their universal application to the study of biological processes in all tissues. Although light can be created by chemiluminescence, light emission from existing chemiluminescent probes is too weak to use this imaging modality in situations when fluorescence cannot be used. Here we report the development of the brightest luminescent protein to date, Nano-lantern, which is a chimera of enhanced Renilla luciferase and Venus, a fluorescent protein with high bioluminescence resonance energy transfer efficiency. Nano-lantern allows real-time imaging of intracellular structures in living cells with spatial resolution equivalent to fluorescence and sensitive tumour detection in freely moving unshaved mice. We also create functional indicators based on Nano-lantern that can image Ca2+, cyclic adenosine monophosphate and adenosine 5′-triphosphate dynamics in environments where the use of fluorescent indicators is not feasible. These luminescent proteins allow visualization of biological phenomena at previously unseen single-cell, organ and whole-body level in animals and plants. Luminescent proteins are important tools for biomedical imaging but tend to emit fairly little light. Saito et al.. describe a brighter version of a bioluminescent protein that can visualize intracellular dynamics of various signalling molecules with high spatial and temporal resolution.
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