On/off blinking and switching behaviour of single molecules of green fluorescent protein
On/off blinking and switching behaviour of single molecules of green fluorescent protein
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DOI：
10.1038/41048
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发表时间：
1997-07-24
期刊：
影响因子：
64.8
通讯作者：
Moerner, WE
中科院分区：
文献类型：
--
作者：
Dickson, RM;Cubitt, AB;Moerner, WE
在低温和室温下对单个分子的光学研究可以提供有关固体、液体和生物系统中局部环境动力学的信息，这些信息不会被系综平均所掩盖（1-14）。在这里，我们提出了一个单分子的绿色荧光蛋白（GFP）的生物物理行为的研究，来自维多利亚水母。野生型GFP及其突变体作为荧光生物标记物引起了人们的兴趣，因为荧光团可以在体内形成（15，16）。GFP突变体固定在aereated水性聚合物凝胶中，并由488 nm光激发，在几秒的时间尺度上经历重复的荧光发射周期（“闪烁”），这在批量研究中是无法观察到的。最终，单个GFP分子达到持久的黑暗状态，通过405 nm的照射，它们可以从黑暗状态切换回原始的发射状态。这表明使用这些GFP作为时间依赖性细胞过程的荧光标记物，以及作为分子光子开关或光学存储元件，在单分子水平上可寻址的可能性。
Optical studies of individual molecules at low and room temperature can provide information about the dynamics of local environments in solids, liquids and biological systems unobscured by ensemble averaging(1-14). Here we present a study of the photophysical behaviour of single molecules of the green fluorescent protein (GFP) derived from the jellfish Aequorea victoria. Wild-type GFP and its mutant have attracted interest as fluorescent biological labels because the fluorophore may be formed in vivo(15,16). GFP mutants immobilized in aereated aqueous polymer gels and excited by 488-nm light undergo repeated cycles of fluorescent emission ('blinking') on a timescale of several seconds-behaviour that would be unobservable in bulk studies. Eventually the individual GFP molecules reach a long-lasting dark state, from which they can be switched back to the original emissive state by irradiation at 405 nm. This suggests the possibility of using these GFPs as fluorescent markers for time-dependent cell processes, and as molecular photonic switches or optical storage elements, addressable on the single-molecule level.