Hohlbein Johannes, Hübner Christian G
Max Planck Institute of Microstructure Physics, Weinberg 2, 06120 Halle, Germany.
J Chem Phys. 2008 Sep 7;129(9):094703. doi: 10.1063/1.2971183.
The power of three-dimensional orientation detection of single emitting dipoles using a sophisticated scheme with three detectors in a confocal microscope is quantitatively explored by means of Monte Carlo simulations. We show that several hundreds of photons are sufficient for a reliable orientation determination. In typical single-molecule experiments, time resolutions in the submillisecond range for orientation trajectories become accessible. Experimental data on fluorescent latex beads and single perylene monoimide molecules show that a properly aligned setup can perfectly reproduce the simulated data. The simulations and experimental data highlight the potential of our method and give practical guidelines for its application.
利用共聚焦显微镜中带有三个探测器的精密方案对单个发射偶极子进行三维方向检测的能力，通过蒙特卡罗模拟进行了定量研究。我们表明，几百个光子就足以进行可靠的方向测定。在典型的单分子实验中，可以实现亚毫秒范围内方向轨迹的时间分辨率。关于荧光乳胶珠和单个苝单酰亚胺分子的实验数据表明，一个正确对齐的装置能够完美地重现模拟数据。模拟和实验数据突出了我们方法的潜力，并为其应用提供了实用指南。
