Title
Stochastic approach to the molecular counting problem in superresolution microscopy
Date Issued
13 January 2015
Access level
open access
Resource Type
journal article
Author(s)
University of California
Publisher(s)
National Academy of Sciences
Abstract
Superresolution imaging methods-now widely used to characterize biological structures below the diffraction limit-are poised to reveal in quantitative detail the stoichiometry of protein complexes in living cells. In practice, the photophysical properties of the fluorophores used as tags in superresolution methods have posed a severe theoretical challenge toward achieving this goal. Here we develop a stochastic approach to enumerate fluorophores in a diffraction-limited area measured by superresolution microscopy. The method is a generalization of aggregated Markov methods developed in the ion channel literature for studying gating dynamics. We show that the method accurately and precisely enumerates fluorophores in simulated data while simultaneously determining the kinetic rates that govern the stochastic photophysics of the fluorophores to improve the prediction's accuracy. This stochastic method overcomes several critical limitations of temporal thresholding methods.
Start page
E110
End page
E118
Volume
112
Issue
2
Language
English
OCDE Knowledge area
Física atómica, molecular y química
Biología celular, Microbiología
Scopus EID
2-s2.0-84920972458
PubMed ID
Source
Proceedings of the National Academy of Sciences of the United States of America
ISSN of the container
00278424
Sponsor(s)
Burroughs Wellcome Fund
Howard Hughes Medical Institute
National Institutes of Health R01-GM032543
National Stroke Foundation MCB-1412259
National Institute of General Medical Sciences T32GM008284 NIGMS
Sources of information:
Directorio de Producción Científica
Scopus