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. 2019 Mar 20;10:1268. doi: 10.1038/s41467-019-09064-6

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — Artifact removal for uniform and quantitative cellular DNA-PAINT imaging. a Full camera chip (130 × 130 µm²) DNA-PAINT image of the microtubule network in fixed COS-7 cells acquired using Gauss illumination (left) and the same field of view for flat-top illumination (right). b Magnified sections from segment 1 and segment 5 (as defined in Fig. 2a) highlighting the image quality in the center and the border region of the camera chip. White arrows point to artifacts due to multi-emitter mislocalizations. c Photon count histograms for box regions in images from b. Double Gaussian fit allows identification and removal of multi-emitter mislocalizations (threshold at 1/e² of first peak) except for segment 5 for Gaussian illumination. d Filtered flat-top images from b displaying enhanced image quality after removing mislocalization artifacts. e Intensity profiles across single microtubules indicated in d. Scale bars, 10 µm in a, 500 nm in b, d