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. 2017 Oct 2;7:12482. doi: 10.1038/s41598-017-12864-9

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© The Author(s) 2017

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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(a) Typical single ThT molecule image depicting the integration area used for the intensity time trace analysis. Scale bar: 500 nm. (b) Typical time trace (backround is removed) obtained at the location of a single ThT molecule (in MEA conditions), showing the standard deviation of the noise $σ$ , and the magnitude of a few $σ$ . (c) Average duty cycle obtained for the different conditions: ThT in MEA (n = 191), ThT in AA (n = 191), Alexa Fluor 488 in MEA (n = 155) deposited on a glass substrate (n: number of time traces over 6 different measurements, for each condition). (d) on-event signal (in camera counts) obtained for the different conditions: ThT in MEA (n = 774), ThT in AA (n = 774), and Alexa Fluor 488 in MEA (n = 774) (n: number of detected on-events, for each condition). The thresholding is $5 σ$ . (e) Photostability duration of detected molecules (e.g. decrease of on-events over time measured from time traces), normalized to the total number of on-events and represented in a cumulative plot, averaged over 6 measurements per condition. (f) Localization precision deduced per detected single molecule in a dSTORM movie stack, for ThT in MEA (n = 1000), ThT in AA (n = 1000), and Alexa Fluor 488 in MEA (n = 1000). All error bars represent the standard deviation over n, number of measured samples.