稀释的自由扩散膜结合荧光团之间稳态Forster能量转移的Stern-Volmer模型
Stern-Volmer Modeling of Steady-State Forster Energy Transfer Between Dilute, Freely Diffusing Membrane-Bound Fluorophores
摘要：
使用两种不同的度量来评估稳态下荧光团之间的福斯特共振能量转移（FRET）：（1）供体荧光的受体猝灭，E（a.k.a.转移效率）;和（ii）供体-激发受体荧光，F-A-Dex.虽然E仍然被更广泛地使用，但F-A-Dex由于实际原因在研究生物膜的实验学家中越来越受欢迎。在这里，对于膜结合的荧光团的特殊情况下，我们提出了大量的实验证据，证明使用简单的斯特恩-沃尔默表达式建模时，在稀释探针条件下的FRET度量。我们还发现了我们的Stern-Volmer表达式E和Wolber和Hudson的系列近似的稳态福斯特淬火在2D之间的稀释制度的对应关系。这种新颖的对应关系使我们能够解释我们的每个二维猝灭常数在两个方面（i）有效的福斯特距离，和（ii）两个最大受体浓度限制，其中每个定义自己的有用的实验制度。综上所述，我们的研究结果提出了一个三步策略，设计更有效的稳态FRET实验的生物膜的研究。
英文摘要：
Two different metrics are used to assess Forster resonance energy transfer (FRET) between fluorophores in the steady state: (1) acceptor-quenching of donor fluorescence, E (a.k.a. transfer efficiency); and (ii) donor-excited acceptor fluorescence, F-A-Dex. While E is still more widely used, F-A-Dex has been gaining in popularity for practical reasons among experimentalists who study biomembranes. Here, for the special case of membrane-bound fluorophores, we present a substantial body of experimental evidence that justifies the use of simple Stern-Volmer expressions when modeling either FRET metric under dilute-probe conditions. We have also discovered a dilute-regime correspondence between our Stern-Volmer expression for E and Wolber and Hudson's series approximation for steady-state Forster quenching in 2D. This novel correspondence allows us to interpret each of our 2D quenching constants in terms of both (i) an effective Forster distance, and (ii) two maximum acceptor-concentration limits, each of which defines its own useful experimental regime. Taken together, our results suggest a three-step strategy toward designing more effective steady-state FRET experiments for the study of biomembranes.
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