Three-dimensional orientation determination of the emission dipoles of single molecules: the shot-noise limit.
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.
Hohlbein
Johannes
J
Max Planck Institute of Microstructure Physics, Weinberg 2, 06120 Halle, Germany. j.hohlbein1@physics.ox.ac.uk
Hübner
Christian G
CG
D005453:Fluorescence; D007094:Imides / Q000737:chemistry*; D007840:Latex / Q000737:chemistry*; D018613:Microscopy, Confocal; D008863:Microspheres; D009010:Monte Carlo Method; D010569:Perylene / Q000031:analogs & derivatives* / Q000737:chemistry; D017785:Photons
D007094:Imides; D007840:Latex; C532868:perylenemonoimide; D010569:Perylene