K-factor image deshadowing for three-dimensional fluorescence microscopy.
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.
Faculty of Engineering, Bar-Ilan University, Ramat-Gan 52900, Israel.
Tali
T
Ilovitsh
Faculty of Engineering, Bar-Ilan University, Ramat-Gan 52900, Israel.
Aryeh
A
Weiss
Department of Electrical and Computer Engineering, University of Utah, Salt Lake City, Utah, USA.
Amihai
A
Meiri
Department of Physics and Astronomy, University of Utah, Salt Lake City, UT.
Carl G
CG
Ebeling
Genetic Institute, Meir Medical Center, Kfar Saba, Israel.|Faculty of Life Sciences, Bar Ilan University, Ramat Gan, Israel.
Aliza
A
Amiel
Genetic Institute, Meir Medical Center, Kfar Saba, Israel.|Faculty of Life Sciences, Bar Ilan University, Ramat Gan, Israel.
Hila
H
Katz
Genetic Institute, Meir Medical Center, Kfar Saba, Israel.|Faculty of Life Sciences, Bar Ilan University, Ramat Gan, Israel.
Batya
B
Mannasse-Green
Faculty of Engineering, Bar-Ilan University, Ramat-Gan 52900, Israel.
Zeev
Z
Zalevsky
D000465:Algorithms; D058948:Cell Tracking / Q000379:methods*; D007089:Image Enhancement / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D010363:Pattern Recognition, Automated / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity