Maximizing the field of view and accuracy in 3D Single Molecule Localization Microscopy.
Optical Society of America (OSA)
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Rehman, S. A., Carr, A., Lenz, M., Lee, S., & O'Holleran, K. (2018). Maximizing the field of view and accuracy in 3D Single Molecule Localization Microscopy.. Optics express, 26 (4), 4631-4637. https://doi.org/10.1364/oe.26.004631
Super-resolution techniques that localize single molecules in three dimensions through point spread function (PSF) engineering are very sensitive to aberrations and optical alignment. Here we show how double-helix point spread function is affected by such mis-alignment and aberration. Specifically, we demonstrate through simulation and experiment how misplacement of phase masks in infinity corrected systems is a common source of significant loss of accuracy. We also describe an optimal alignment and calibration procedure to correct for these errors. In combination, these optimizations allow for a maximal field of view with high accuracy and precision. Though discussed with reference to double-helix point spread function (DHPSF), the optimization techniques are equally applicable to other engineered PSFs.
UNIVERSITY COLLEGE LONDON (FB EPSRC) (EP/L015455/1)
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External DOI: https://doi.org/10.1364/oe.26.004631
This record's URL: https://www.repository.cam.ac.uk/handle/1810/274123
Attribution 4.0 International
Licence URL: http://creativecommons.org/licenses/by/4.0/
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