化学性质相同的相邻荧光团的非加性效应模型
Modeling Non-additive Effects in Neighboring Chemically Identical Fluorophores
摘要：
定量荧光分析通常用于推导生物分子复合物的化学性质，包括化学计量。在荧光数据分析中的一个基本假设--无论是通过超分辨率测定蛋白质复合物的化学计量，还是通过光漂白进行步进计数，或者在RNA荧光{ \it原位}杂交（RNA-FISH）实验中测定衍射限制点中的RNA计数--是荧光团的行为相同并且不相互作用。然而，最近对荧光团标记的DNA折纸结构（如荧光立方体）的实验揭示了相同荧光团之间相互作用的性质，因为这些荧光团更靠近在一起，从而对荧光团不相互作用的建模假设的有效性提出了质疑。在这里，我们分析脉冲照明下的光子到达数据从荧光立方体染料之间的距离范围从2-10 nm。我们讨论了亮度的非加和性对定量荧光分析的影响。
英文摘要：
Quantitative fluorescence analysis is often used to derive chemical properties, including stoichiometries, of biomolecular complexes. One fundamental underlying assumption in the analysis of fluorescence data -- whether it be the determination of protein complex stoichiometry by super-resolution, or step-counting by photobleaching, or the determination of RNA counts in diffraction-limited spots in RNA fluorescence {\it in situ} hybridization (RNA-FISH) experiments -- is that fluorophores behave identically and do not interact. However, recent experiments on fluorophore-labeled DNA-origami structures such as fluorocubes have shed light on the nature of the interactions between identical fluorophores as these are brought closer together, thereby raising questions on the validity of the modeling assumption that fluorophores do not interact. Here, we analyze photon arrival data under pulsed illumination from fluorocubes where distances between dyes range from 2-10 nm. We discuss the implications of non-additivity of brightness on quantitative fluorescence analysis.
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