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

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 t...

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Veröffentlicht in:Optics express 2014-05, Vol.22 (9), p.10304-10316
Hauptverfasser: Proppert, Sven, Wolter, Steve, Holm, Thorge, Klein, Teresa, van de Linde, Sebastian, Sauer, Markus
Format: Artikel
Sprache:eng
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Zusammenfassung: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 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.
ISSN:1094-4087
1094-4087
DOI:10.1364/OE.22.010304