Quantifying Microsecond Transition Times Using Fluorescence Lifetime Correlation Spectroscopy
Quantifying Microsecond Transition Times Using Fluorescence Lifetime Correlation Spectroscopy
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DOI：
10.1021/acs.jpclett.7b02707
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发表时间：
2017-12-06
影响因子：
5.7
通讯作者：
Karedla, Narain
中科院分区：
文献类型：
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作者：
Ghosh, Arindam;Isbaner, Sebastian;Karedla, Narain
许多复杂的发光体，如荧光蛋白，表现出多种发射状态，导致其激发态寿命的快速波动。本文应用荧光寿命相关光谱（FLCS）分析了典型荧光蛋白增强型绿色荧光蛋白（EGFP）的光物理状态动力学。我们量化了其两个荧光状态之间的微秒跃迁率，这两个荧光状态具有高度重叠的发射光谱。我们将这些转变与室温下氨基酸在荧光中心附近的埃级旋转异构体联系起来。通过这项研究，我们证明了FLCS在研究具有复杂多态光物理的广泛发光系统的快速跃迁动力学方面的能力，这是其他方法无法轻易完成的。
Many complex luminescent emitters such as fluorescent proteins exhibit multiple emitting states that result in rapid fluctuations of their excited-state lifetime. Here, we apply fluorescence lifetime correlation spectroscopy (FLCS) to resolve the photophysical state dynamics of the prototypical fluorescence protein enhanced green fluorescent protein (EGFP). We quantify the microsecond transition rates between its two fluorescent states, which have otherwise highly overlapping emission spectra. We relate these transitions to a room temperature angstrom-scale rotational isomerism of an amino acid next to its fluorescent center. With this study, we demonstrate the power of FLCS for studying the rapid transition dynamics of a broad range of light-emitting systems with complex multistate photophysics, which cannot be easily done by other methods.