K-factor image deshadowing for three-dimensional fluorescence microscopy

Sci Rep. 2015 Sep 3:5:13724. doi: 10.1038/srep13724.

Abstract

The ability to track single fluorescent particles within a three dimensional (3D) cellular environment can provide valuable insights into cellular processes. In this paper, we present a modified nonlinear image decomposition technique called K-factor that reshapes the 3D point spread function (PSF) of an XYZ image stack into a narrow Gaussian profile. The method increases localization accuracy by ~60% with compare to regular Gaussian fitting, and improves minimal resolvable distance between overlapping PSFs by ~50%. The algorithm was tested both on simulated data and experimentally.

Publication types

  • Evaluation Study

MeSH terms

  • Algorithms*
  • Cell Tracking / methods*
  • Image Enhancement / methods
  • Image Interpretation, Computer-Assisted / methods*
  • Imaging, Three-Dimensional / methods*
  • Microscopy, Fluorescence / methods*
  • Pattern Recognition, Automated / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity