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Measuring localization performance of super-resolution algorithms on very active samples

Please always quote using this URN: urn:nbn:de:bvb:20-opus-85936
  • Super-resolution fluorescence imaging based on inglemolecule localization relies critically on the availability of efficient processing algorithms to distinguish, identify, and localize emissions of single fluorophores. In multiple current applications, such as threedimensional, time-resolved or cluster imaging, high densities of fluorophore emissions are common. Here, we provide an analytic tool to test the performance and quality of localization microscopy algorithms and demonstrate that common algorithms encounter difficulties for samplesSuper-resolution fluorescence imaging based on inglemolecule localization relies critically on the availability of efficient processing algorithms to distinguish, identify, and localize emissions of single fluorophores. In multiple current applications, such as threedimensional, time-resolved or cluster imaging, high densities of fluorophore emissions are common. Here, we provide an analytic tool to test the performance and quality of localization microscopy algorithms and demonstrate that common algorithms encounter difficulties for samples with high fluorophore density. We demonstrate that, for typical single-molecule localization microscopy methods such as dSTORM and the commonly used rapidSTORM scheme, computational precision limits the acceptable density of concurrently active fluorophores to 0.6 per square micrometer and that the number of successfully localized fluorophores per frame is limited to 0.2 per square micrometer.show moreshow less

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Metadaten
Author: Steve Wolter, Ulrike Endesfelder, Sebastian van de Linde, Mike Heilemann, Markus Sauer
URN:urn:nbn:de:bvb:20-opus-85936
Document Type:Journal article
Faculties:Fakultät für Physik und Astronomie / Physikalisches Institut
Language:English
Parent Title (English):Optics Express
Year of Completion:2011
Source:Optics Express (2011) 19:8, 7020-7033, doi:10.1364/OE.19.007020
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 53 Physik / 530 Physik
Release Date:2014/05/12
Collections:Open-Access-Publikationsfonds / Förderzeitraum 2011
Licence (German):License LogoDeutsches Urheberrecht