Localization microscopy at doubled precision with patterned illumination
Jelmer Cnossen (TU Delft - ImPhys/Quantitative Imaging, TU Delft - Mechanical Engineering)
Taylor Hinsdale (TU Delft - ImPhys/Quantitative Imaging)
Rasmus Thorsen (TU Delft - ImPhys/Quantitative Imaging)
Marijn Siemons (Universiteit Utrecht)
Florian Schueder (Max Planck Institute of Biochemistry, LMU Munich)
Ralf Jungmann (Max Planck Institute of Biochemistry, LMU Munich)
Carlas S. Smith (TU Delft - ImPhys/Quantitative Imaging, TU Delft - Mechanical Engineering, University of Oxford)
Bernd Rieger (TU Delft - ImPhys/Quantitative Imaging)
Sjoerd Stallinga (TU Delft - Applied Sciences)
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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.