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

Sven Proppert, Steve Wolter, Thorge Holm, Teresa Klein, Sebastian van de Linde, Markus Sauer

Research output: Contribution to journalArticlepeer-review

16 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)10304-10316
Number of pages13
JournalOptics Express
Issue number9
Early online date22 Apr 2014
Publication statusPublished - 5 May 2014
Externally publishedYes


  • cubic B-splines
  • 3D super-resolution fluorescence imaging
  • localization microscopy
  • astigmatic imaging


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