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Cubic B-spline calibration for 3D super-resolution measurements using astigmatic imaging

Please always quote using this URN: urn:nbn:de:bvb:20-opus-119730
  • In recent years three-dimensional (3D) super-resolution fluorescence imaging by single-molecule localization (localization microscopy) has gained considerable interest because of its simple implementation and high optical resolution. Astigmatic and biplane imaging are experimentally simple methods to engineer a 3D-specific point spread function (PSF), but existing evaluation methods have proven problematic in practical application. Here we introduce the use of cubic B-splines to model the relationship of axial position and PSF width in theIn recent years three-dimensional (3D) super-resolution fluorescence imaging by single-molecule localization (localization microscopy) has gained considerable interest because of its simple implementation and high optical resolution. Astigmatic and biplane imaging are experimentally simple methods to engineer a 3D-specific point spread function (PSF), but existing evaluation methods have proven problematic in practical application. Here we introduce the use of cubic B-splines to model the relationship of axial position and PSF width in the above mentioned approaches and compare the performance with existing methods. We show that cubic B-splines are the first method that can combine precision, accuracy and simplicity.show moreshow less

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Metadaten
Author: Sven Proppert, Steve Wolter, Thorge Holm, Theresa Klein, Sebastian van de Linde, Markus Sauer
URN:urn:nbn:de:bvb:20-opus-119730
Document Type:Journal article
Faculties:Fakultät für Biologie / Theodor-Boveri-Institut für Biowissenschaften
Language:English
Parent Title (English):Optics Express
ISSN:1094-4087
Year of Completion:2014
Volume:22
Issue:9
Pagenumber:10304-16
Source:Optics Express 22,9 p.10304-16. DOI:10.1364/OE.22.010304
DOI:https://doi.org/10.1364/OE.22.010304
Pubmed Id:http://www.ncbi.nlm.nih.gov/pubmed?term=24921733
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 50 Naturwissenschaften / 502 Verschiedenes
Tag:fluorescence microscopy; medical and biological imaging; superresolution
three-dimensional microscopy
Release Date:2015/11/12
Licence (German):License LogoDeutsches Urheberrecht