Smoothness correction for better SOFI imaging
Siewert Hugelier (Katholieke Universiteit Leuven)
Wim Vandenberg (Katholieke Universiteit Leuven)
Tomáš Lukeš (École Polytechnique Fédérale de Lausanne)
Kristin S. Grußmayer (École Polytechnique Fédérale de Lausanne, TU Delft - BN/Kristin Grussmayer Lab)
Paul H.C. Eilers (Erasmus MC)
Peter Dedecker (Katholieke Universiteit Leuven)
Cyril Ruckebusch (Université de Lille)
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Abstract
Sub-diffraction or super-resolution fluorescence imaging allows the visualization of the cellular morphology and interactions at the nanoscale. Statistical analysis methods such as super-resolution optical fluctuation imaging (SOFI) obtain an improved spatial resolution by analyzing fluorophore blinking but can be perturbed by the presence of non-stationary processes such as photodestruction or fluctuations in the illumination. In this work, we propose to use Whittaker smoothing to remove these smooth signal trends and retain only the information associated to independent blinking of the emitters, thus enhancing the SOFI signals. We find that our method works well to correct photodestruction, especially when it occurs quickly. The resulting images show a much higher contrast, strongly suppressed background and a more detailed visualization of cellular structures. Our method is parameter-free and computationally efficient, and can be readily applied on both two-dimensional and three-dimensional data.
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