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.
Original language | English |
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Article number | 7569 |
Journal | Scientific Reports |
Volume | 11 |
Issue number | 1 |
DOIs | |
Publication status | Published - 7 Apr 2021 |
Bibliographical note
Funding Information:S.H. thanks the Research Foundation Flanders for a postdoctoral fellowship and the French embassy in Belgium for a PHC Tournesol mobility grant. P.D. and W.V. thank the Research Foundation Flanders through Grants 1514319N, G090819N, G0B8817N, and the European Research Council through Grant 714688 NanoCellActiv-ity. C.R. and P.D. acknowledge the LAI High Performance Fluorescence Microscopy from U LILLE–KU Leuven. K. S. G. acknowledges the support from the Horizon 2020 Framework Program of the European Union under the Marie Skłodowska-Curie Grant Agreement No. [750528] and thanks Anne-Laure Mahul-Mellier (LMNN, EPFL) for providing neurons.
Publisher Copyright:
© 2021, The Author(s).