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. 2018 Oct 11;8:15154. doi: 10.1038/s41598-018-33225-0

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© The Author(s) 2018

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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qSMLM results corroborate FISH analysis. (A) A touch prep region with individual localizations (blue) rendered by a Gaussian blur. The intensity of the Gaussian blur is proportional to the localization density (full cell, scale bar: 5 μm; zoomed-in region, scale bar: 100 nm). (B) Average detected HER2 density from qSMLM versus HER2 copy number from FISH. The following number of ROIs were used for analysis: P1 (30 ROIs), P3 (92 ROIs), P4 (88 ROIs), P5 (131 ROIs), P6 (164 ROIs), P7 (46 ROIs). Complete FISH data was not available for P2. The fitted least-square line has a slope estimate of 0.70, and an intercept estimate of −9.63 (R² = 0.958). (C) Fraction of ROIs above HER2 density threshold. (D) The average HER2 receptors per cluster versus HER2 copy number from FISH. Clustered regions were used for analysis (19 ROI for P1, 44 ROI for P3, 28 ROI for P4, 10 ROI for P5, and 14 ROI for P6). The fitted least-square line has a slope estimate of 12.45, and an intercept estimate of −29.75 (R² = 0.88). Error bars are SEM. Additional statistical considerations are included in Table 1.