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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:荧光相关光谱(FCS)已被广泛应用于研究分子的动力学和光学药物以及分子之间的相互作用,通过从信号的波动提取信息。特别是,FCS的单一分子应用承诺是最大的信息。理想情况下,人们想实时进行FCS;然而,由于相关过程的耗时性质,实时执行相关性是完全不变的。通常,通常需要昂贵的硬件相关器或TCSPC板。最近,已经提出了基于多TAU方法的高效算法来构建软件相关器。在这项工作中,我们阐述了一种能够实现实时相关性的创新算法,而无需转向多Tau方法。该算法利用通常存在于FCS实验中的低计数率,直接使用每个光子与其相邻光子之间的时间间隔有效更新相关函数。基于这种效率,可以仅用普通计数器构建低成本的软件相关器。我们实际上通过建立这种相关器来测量罗丹明6g在水中使用FCS来测量水中的可行性。通过将来自光子检测器的信号复制并通过我们的软件相关器和商业相关器同时将信号与普通计数器发送到普通计数器和商业相关器来验证该算法。来自这两个相关器的相关曲线的完美巧合和相关函数的实时显示表明我们方法的有效性和实用性。

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