Resonance energy transfer (RET) and fluorescence fluctuation spectroscopies (FFS) are powerful fluorescence-based techniques for quantifying the self-association of membrane receptors within oligomeric complexes in living cells. However, RET spectrometry’s ability to extract information on the detailed quaternary structure of oligomers sometimes rests on assumptions regarding the relative abundances of oligomers of different sizes, while FFS techniques may provide oligomer size information but not quaternary structure details, as they lack a probe for inter-molecular distances. In this report, we introduce a method which we termed ‘intensity fluctuations and resonance energy transfer’ (IFRET), which combines analysis of donor and acceptor intensity fluctuations with RET efficiency determination. Because the three measured quantities each have a unique dependence on the acceptor mole fraction ( X A ), simultaneous global fitting of all three dramatically ...

中文翻译：

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建议同时分析荧光强度波动和共振能量转移（IFRET）测量。

共振能量转移（RET）和荧光波动光谱（FFS）是基于荧光的强大技术，用于量化活细胞中寡聚复合物中膜受体的自缔合。但是，RET光谱法提取低聚物详细四级结构信息的能力有时取决于关于不同大小低聚物的相对丰度的假设，而FFS技术可能提供低聚物尺寸信息，但不能提供四级结构细节，因为它们缺乏相互间的探针。 -分子距离。在本报告中，我们介绍了一种称为“强度波动和共振能量转移”（IFRET）的方法，该方法将供体和受体强度波动的分析与RET效率确定相结合。