Quantitative assessment of fluorescent proteins.
Quantitative assessment of fluorescent proteins.
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DOI：
10.1038/nmeth.3891
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发表时间：
2016-07
期刊：
影响因子：
48
通讯作者：
Piston DW
中科院分区：
文献类型：
--
作者：
Cranfill PJ;Sell BR;Baird MA;Allen JR;Lavagnino Z;de Gruiter HM;Kremers GJ;Davidson MW;Ustione A;Piston DW
用于分子和细胞遗传标记的荧光蛋白（FP）的出现彻底改变了荧光显微镜。基因操作已经创造了大量明亮而稳定的FP，这些FP跨越了蓝色到红色的光谱区域。自发荧光FP的共同点是它们的紧密β-桶结构，其提供了有效荧光所需的刚性和化学环境。尽管有共同的结构，但每个FP都有自己独特的物理性质。因此，对于每种情况都没有单一的“最佳”荧光蛋白，每种FP都有优点和缺点。为了指导决定哪种FP适合任何给定的应用，我们对40多种不同的FP的亮度、光稳定性、pH稳定性和单体特性进行了定量表征，这使得这些FP之间的比较变得容易。我们报告了所有测量的FP的值，但将讨论集中在每个光谱区域中更流行和/或性能最好的FP上。
The advent of fluorescent proteins (FP) for genetic labeling of molecules and cells has revolutionized fluorescence microscopy. Genetic manipulations have created a vast array of bright and stable FPs spanning the blue to red spectral regions. Common to autofluorescent FPs is their tight β-barrel structure, which provides the rigidity and chemical environment needed for effectual fluorescence. Despite the common structure, each FP has its own unique photophysical properties. Thus, there is no single “best” fluorescent protein for every circumstance, and each FP has advantages and disadvantages. To guide decisions about which FP is right for any given application, we have characterized quantitatively over 40 different FPs for their brightness, photostability, pH stability, and monomeric properties, which permits easy apples-to-apples comparisons between these FPs. We report the values for all of the FPs measured, but focus the discussion on the more popular and/or best performing FPs in each spectral region.