Measuring localization performance of super-resolution algorithms on very active samples
Wolter, Steve and Endesfelder, Ulrike and van de Linde, Sebastian and Heilemann, Mike and Sauer, Markus (2011) Measuring localization performance of super-resolution algorithms on very active samples. Optics Express, 19 (8). pp. 7020-7033. ISSN 1094-4087 (https://doi.org/10.1364/OE.19.007020)
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Abstract
Super-resolution fluorescence imaging based on singlemolecule 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.
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Item type: Article ID code: 77990 Dates: DateEvent29 March 2011Published27 February 2011AcceptedSubjects: Science > Physics > Optics. Light Department: Faculty of Science > Physics Depositing user: Pure Administrator Date deposited: 04 Oct 2021 09:44 Last modified: 11 Nov 2024 13:15 Related URLs: URI: https://strathprints.strath.ac.uk/id/eprint/77990