Real-time adaptive drift correction for super-resolution localization microscopy

Super-resolution localization microscopy involves acquiring thousands of image frames of sparse collections of single molecules in the sample. The long acquisition time makes the imaging setup prone to drift, affecting accuracy and precision. Localization accuracy is generally improved by a posterio...

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Bibliographic Details
Published in:Optics express 2015-09, Vol.23 (18), p.23887-23898
Main Authors: Grover, Ginni, Mohrman, Wyatt, Piestun, Rafael
Format: Article
Language:English
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Summary:Super-resolution localization microscopy involves acquiring thousands of image frames of sparse collections of single molecules in the sample. The long acquisition time makes the imaging setup prone to drift, affecting accuracy and precision. Localization accuracy is generally improved by a posteriori drift correction. However, localization precision lost due to sample drifting out of focus cannot be recovered as the signal is originally detected at a lower peak signal. Here, we demonstrate a method of stabilizing a super-resolution localization microscope in three dimensions for extended periods of time with nanometer precision. Hence, no localization correction after the experiment is required to obtain super-resolved reconstructions. The method incorporates a closed-loop with a feedback signal generated from camera images and actuation on a 3D nanopositioning stage holding the sample.
ISSN:1094-4087
1094-4087
DOI:10.1364/OE.23.023887