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High-speed low-cost correlator for single molecule fluorescence correlation spectroscopy

机译:用于单分子荧光相关光谱的高速低成本相关器

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Fluorescence correlation spectroscopy (FCS) has been extensively applied to study the kinetics and photophysics of molecules as well as interactions between molecules by extracting information from the fluctuation of signals. In particular, single molecule applications of FCS promise the greatest amounts of information. Ideally, one would like to carry out FCS in real-time; however, due to the time-consuming nature of the correlation process, performing the correlation in real-time is totally nontrivial. Generally an expensive hardware correlator or a TCSPC board is required for this purpose. Recently highly-efficient algorithms based on multi-tau method have been proposed to build up a software correlator. In this work, we set forth an innovative algorithm capable of realizing the real-time correlation, without turning to the multi-tau method. This algorithm takes advantage of the low count rate generally existing in the FCS experiments, directly using the time interval between each photon its adjacent photon to efficiently update the correlation function. Based on this efficiency, it is possible to build a low-cost software correlator with just an ordinary counter board. We practically demonstrate the feasibility by setting up this correlator to measure the diffusion motion of rhodamine 6G in water using FCS. The algorithm was validated by duplicating the signal from the photon detector and sending it to both the ordinary counter board with our software correlator and a commercial correlator simultaneously. The perfect coincidence of the correlation curves from these two correlators and the real-time display of the correlation function indicate the validity and practicability of our approach.
机译:通过从信号波动中提取信息,荧光相关光谱(FCS)已被广泛应用于研究分子的动力学和光物理以及分子之间的相互作用。特别是,FCS的单分子应用有望提供最多的信息。理想情况下,希望实时进行FCS;但是,由于关联过程很耗时,因此实时进行关联是非常重要的。通常,为此目的需要昂贵的硬件相关器或TCSPC板。最近提出了一种基于多τ方法的高效算法来构建软件相关器。在这项工作中,我们提出了一种创新的算法,该算法能够实现实时相关,而无需转向多tau方法。该算法直接利用FCS实验中通常存在的低计数率,直接使用每个光子与其相邻光子之间的时间间隔来有效地更新相关函数。基于这种效率,有可能仅使用普通的计数器板就可以构建低成本的软件相关器。我们通过设置此相关器以使用FCS测量若丹明6G在水中的扩散运动来实际证明其可行性。通过复制来自光子检测器的信号并同时使用我们的软件相关器和商用相关器将其发送到普通计数器板来验证算法。来自这两个相关器的相关曲线的完美重合以及相关函数的实时显示表明了我们方法的有效性和实用性。

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