Localization microscopy at doubled precision with patterned illumination
Jelmer Cnossen (TU Delft - ImPhys/Quantitative Imaging, TU Delft - Team Raf Van de Plas)
Taylor Hindsdale (TU Delft - ImPhys/Quantitative Imaging)
R. O. Thorsen (TU Delft - ImPhys/Quantitative Imaging)
Marijn Siemons (Universiteit Utrecht)
Florian Schueder (Ludwig Maximilians University, Max Planck Institute of Biochemistry)
Ralf Jungmann (Max Planck Institute of Biochemistry, Ludwig Maximilians University)
C.S. Smith (TU Delft - Team Raf Van de Plas, University of Oxford, TU Delft - ImPhys/Quantitative Imaging)
B Rieger (TU Delft - ImPhys/Quantitative Imaging)
S. Stallinga (TU Delft - ImPhys/Imaging Physics)
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Abstract
MINFLUX offers a breakthrough in single molecule localization precision, but is limited in field of view. Here we combine centroid estimation and illumination pattern induced photon count variations in a conventional widefield imaging setup to extract position information over a typical micrometer-sized field of view. We show a near two-fold improvement in precision over standard localization with the same photon count on DNA-origami nanostructures and tubulin in cells, using DNA-PAINT and STORM imaging.