Optimized localization analysis for single-molecule tracking and super-resolution microscopy

Nat Methods. 2010 May;7(5):377-81. doi: 10.1038/nmeth.1447. Epub 2010 Apr 4.

Abstract

We optimally localized isolated fluorescent beads and molecules imaged as diffraction-limited spots, determined the orientation of molecules and present reliable formulas for the precision of various localization methods. Both theory and experimental data showed that unweighted least-squares fitting of a Gaussian squanders one-third of the available information, a popular formula for its precision exaggerates beyond Fisher's information limit, and weighted least-squares may do worse, whereas maximum-likelihood fitting is practically optimal.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Image Interpretation, Computer-Assisted / methods*
  • Likelihood Functions
  • Microscopy, Fluorescence / methods*
  • Nanotechnology
  • Photons