Department of Physics and Astronomy, Faculty of Sciences, VU University Amsterdam, De Boelelaan 1081, 1081 HV Amsterdam, The Netherlands.
J Phys Chem B. 2011 May 12;115(18):5071-82. doi: 10.1021/jp201609c. Epub 2011 Mar 31.
We present a simple method to resolve discrete intensity shifts from time-resolved single-molecule emission data. This new method uses multiples of the standard deviation of the measured intensities that are integrated into short time bins. By applying the technique to trimeric units of the main light-harvesting complex (LHCII) of plants, it is shown that the amount of information that can be extracted from an intensity time trace increases considerably, thereby enlarging the possibility to reveal new phenomena. It is demonstrated how shot noise can lead to substantial deviations and misleading interpretations when the conventional two-state kinetic model for intensity fluctuations is applied. By first resolving the accessed intensity levels, the artifactual effect of shot noise is sufficiently reduced. The technique is particularly applicable to the analysis of fluorescence intermittency from multichromophoric systems.
我们提出了一种简单的方法来解决时间分辨单分子发射数据中的离散强度偏移问题。这种新方法使用多个标准偏差来测量积分到短时间窗口的强度。通过将该技术应用于植物主要光捕获复合物（LHCII）的三聚体单元，结果表明可以从强度时间轨迹中提取的信息量大大增加，从而增加了揭示新现象的可能性。结果表明，当应用传统的强度波动双态动力学模型时，散粒噪声会导致显著的偏差和误导性的解释。通过首先解析可访问的强度水平，可以充分降低散粒噪声的人为影响。该技术特别适用于多色体系荧光间歇性的分析。
