Journal
OPTICS EXPRESS
Volume 23, Issue 18, Pages 23887-23898Publisher
OPTICAL SOC AMER
DOI: 10.1364/OE.23.023887
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Funding
- NSF [1063407, 1429782, 0801680]
- Direct For Biological Sciences
- Div Of Biological Infrastructure [1063407, 1429782] Funding Source: National Science Foundation
- Direct For Education and Human Resources
- Division Of Graduate Education [0801680] Funding Source: National Science Foundation
- Directorate For Engineering
- Div Of Industrial Innovation & Partnersh [1353638] Funding Source: National Science Foundation
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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. (C) 2015 Optical Society of America
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