Localizing and tracking single nanoscale emitters in three dimensions with high spatiotemporal resolution using a double-helix point spread function.
Three-dimensional nanoscale localization and tracking of dim single emitters can be obtained with a widefield fluorescence microscope exhibiting a double-helix point spread function (DH-PSF). We describe in detail how the localization precision quantitatively depends upon the number of photons detected and the z position of the nanoscale emitter, thereby showing a approximately 10 nm localization capability along x, y, and z in the limit of weak emitters. Experimental measurements are compared to Fisher information calculations of the ultimate localization precision inherent in the DH-PSF. The DH-PSF, for the first time, is used to track single quantum dots in aqueous solution and a quantum dot-labeled structure inside a living cell in three dimensions.
Thompson
Michael A
MA
Department of Chemistry, Stanford University, Stanford, CA 94305, USA.
Lew
Matthew D
MD
Badieirostami
Majid
M
Moerner
W E
WE
D045744:Cell Line, Tumor; D004867:Equipment Design; D006801:Humans; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D015233:Models, Statistical; D053758:Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods; D016011:Normal Distribution; D017785:Photons; D045663:Quantum Dots