Institut für Physikalische Chemie, Universität Mainz, D-55099 Mainz, Germany.
J Chem Phys. 2010 Jan 28;132(4):044509. doi: 10.1063/1.3303634.
We depict two algorithms to calculate correlation functions from two different time resolved single molecule fluorescence experiments without the need of time binning. Our first procedure allows to calculate the reduced linear dichroism from polarization resolved fluorescence data. Since we process single photon counts instead of time binned data, considerably faster fluctuations of the dichroism can be analyzed than with conventional methods. With our second procedure time resolved fluorescence obtained with a time correlated single photon counting setup can be analyzed with respect to fluorescence lifetime fluctuations. Again this new algorithm processes single photon events making time binning of photon counts obsolete. Both methods presented are characterized by enhanced time resolution thus allowing to study fast fluctuations of either single molecular orientation or fluorescence life times, respectively.
我们描述了两种算法,无需时间-bin 即可从两个不同的时间分辨单分子荧光实验中计算相关函数。我们的第一个程序允许从偏振分辨荧光数据中计算简化的线二向色性。由于我们处理的是单光子计数而不是时间 bin 数据,因此可以比传统方法更快地分析二向色性的波动。我们的第二个程序可以分析使用时间相关单光子计数设置获得的时间分辨荧光相对于荧光寿命波动。同样,这个新算法处理单光子事件,使得光子计数的时间 bin 变得多余。所提出的两种方法的特点是增强的时间分辨率,从而允许分别研究单分子取向或荧光寿命的快速波动。