title: Accounting for limited detection efficiency and localization precision in cluster analysis in single molecule localization microscopy.
abstract: Single Molecule Localization Microscopy techniques like PhotoActivated Localization Microscopy, with their sub-diffraction limit spatial resolution, have been popularly used to characterize the spatial organization of membrane proteins, by means of quantitative cluster analysis. However, such quantitative studies remain challenged by the techniques' inherent sources of errors such as a limited detection efficiency of less than 60%, due to incomplete photo-conversion, and a limited localization precision in the range of 10-30 nm, varying across the detected molecules, mainly depending on the number of photons collected from each. We provide analytical methods to estimate the effect of these errors in cluster analysis and to correct for them. These methods, based on the Ripley's L(r) - r or Pair Correlation Function popularly used by the community, can facilitate potentially breakthrough results in quantitative biology by providing a more accurate and precise quantification of protein spatial organization.
authors: Laboratory of Nanoscale Biology, Institute of Bioengineering, Ecole Polytechnique Federale de Lausanne (EPFL), Lausanne, Switzerland.
authors: Arun
authors: A
authors: Shivanandan
authors: Audiovisual Communications Laboratory, School of Computer and Communication Sciences, Ecole Polytechnique Federale de Lausanne (EPFL), Lausanne, Switzerland.
authors: Jayakrishnan
authors: J
authors: Unnikrishnan
authors: Laboratory of Nanoscale Biology, Institute of Bioengineering, Ecole Polytechnique Federale de Lausanne (EPFL), Lausanne, Switzerland.
authors: Aleksandra
authors: A
authors: Radenovic
mesh_terms: D016000:Cluster Analysis; D003198:Computer Simulation; D007091:Image Processing, Computer-Assisted; D057230:Limit of Detection; D008853:Microscopy / Q000379:methods*