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Bayesian localization microscopy reveals nanoscale podosome dynamics.

ABSTRACT: We describe a localization microscopy analysis method that is able to extract results in live cells using standard fluorescent proteins and xenon arc lamp illumination. Our Bayesian analysis of the blinking and bleaching (3B analysis) method models the entire dataset simultaneously as being generated by a number of fluorophores that may or may not be emitting light at any given time. The resulting technique allows many overlapping fluorophores in each frame and unifies the analysis of the localization from blinking and bleaching events. By modeling the entire dataset, we were able to use each reappearance of a fluorophore to improve the localization accuracy. The high performance of this technique allowed us to reveal the nanoscale dynamics of podosome formation and dissociation throughout an entire cell with a resolution of 50 nm on a 4-s timescale.

SUBMITTER: Cox S

PROVIDER: S-EPMC3272474 | biostudies-literature | 2011 Dec

REPOSITORIES: biostudies-literature

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Bayesian localization microscopy reveals nanoscale podosome dynamics.

Cox Susan S Rosten Edward E Monypenny James J Jovanovic-Talisman Tijana T Burnette Dylan T DT Lippincott-Schwartz Jennifer J Jones Gareth E GE Heintzmann Rainer R

Nature methods 20111204 2

We describe a localization microscopy analysis method that is able to extract results in live cells using standard fluorescent proteins and xenon arc lamp illumination. Our Bayesian analysis of the blinking and bleaching (3B analysis) method models the entire dataset simultaneously as being generated by a number of fluorophores that may or may not be emitting light at any given time. The resulting technique allows many overlapping fluorophores in each frame and unifies the analysis of the locali ...[more]

PMID: 22138825

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