Spectrally resolved frequency domain analysis of multi-fluorophore systems undergoing energy transfer
Spectrally resolved frequency domain analysis of multi-fluorophore systems undergoing energy transfer
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DOI：
10.1366/000370206779321544
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发表时间：
2006-12-01
影响因子：
3.5
通讯作者：
Hanley, Quentin S.
中科院分区：
文献类型：
--
作者：
Forde, Toni S.;Hanley, Quentin S.
当用频域方法探测时，经历能量转移的荧光团的复杂系统可以表现出各种异常的寿命行为。当以传统的表观寿命格式呈现时，来自这样的系统的数据可以表现出“节点”行为，其中计算的寿命接近+/-无穷大。节点的位置取决于系统和频率。此外，由于典型测量中的噪声，不经历能量转移的较简单系统在零寿命（in）和长寿命（tau（phi））区域中显示出不良行为。在这里，我们系统地调查系统的多个荧光团和没有能量转移提供深入了解频域调查的复杂系统的荧光团。模拟的结果进行比较，从多荧光团系统收集的数据，旨在表现出荧光共振能量转移（FRET）使用成像光谱荧光寿命成像显微镜（ISFLIM）。结果适用于比色皿和成像装置。
Complex systems of fluorophores undergoing energy transfer can exhibit a variety of anomalous lifetime behavior when probed with frequency domain methods. When presented in traditional apparent lifetime format the data from such systems can exhibit "nodal" behavior in which the computed lifetime approaches +/-infinity. The location of the nodes is system and frequency dependent. In addition, simpler systems, not undergoing energy transfer, show ill behavior in the region of zero lifetime (in) and long lifetime (tau(phi)) due to noise in typical measurements. Here, we systematically investigate systems of multiple fluorophores with and without energy transfer to provide insight into frequency domain investigations of complex systems of fluorophores. The results of simulations are compared to data collected from a multi-fluorophore system designed to exhibit fluorescence resonance energy transfer (FRET) using imaging spectroscopic fluorescence lifetime imaging microscopy (ISFLIM). The results are applicable to both cuvette and imaging arrangements.