Accurate construction of photoactivated localization microscopy (PALM) images for quantitative measurements.

ABSTRACT: Localization-based superresolution microscopy techniques such as Photoactivated Localization Microscopy (PALM) and Stochastic Optical Reconstruction Microscopy (STORM) have allowed investigations of cellular structures with unprecedented optical resolutions. One major obstacle to interpreting superresolution images, however, is the overcounting of molecule numbers caused by fluorophore photoblinking. Using both experimental and simulated images, we determined the effects of photoblinking on the accurate reconstruction of superresolution images and on quantitative measurements of structural dimension and molecule density made from those images. We found that structural dimension and relative density measurements can be made reliably from images that contain photoblinking-related overcountin

SUBMITTER: Coltharp C 

PROVIDER: S-EPMC3520911 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature