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. 2017 Dec 20;7:17957. doi: 10.1038/s41598-017-18303-z

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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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Congested arrays. In congested images, the number of single molecules is estimated by dividing the total array spot intensity by the average single molecule intensity, which is estimated using the single molecule algorithms described above on non-congested images. Results show the performance of the algorithms to data with a signal molecule signal to noise ratio of (a) 5, (b) 7.5 (solid lines) and 10 (dashed lines). The figure labels ‘No Filter’, ‘A Trous Wavelet’ and ‘Ricker Wavelet’ represent the results when using estimates of average single molecule intensity determined using the intensity thresholding algorithms with either no filters or pre-processing, or pre-processing with the ‘à trous’ or Ricker wavelet transforms, respectively. Similarly, the ‘PeakFit’ figure label represents the results when using an estimate of average single molecule intensity determined using the peak fitting algorithm.