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. 2022 Mar 18;12:4682. doi: 10.1038/s41598-022-08245-6

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Effect of photobleaching and labeling efficiency on unhindered growth and restricted growth that stops after the assembly of four monomers. Both unhindered (A–D) and restricted growth (E–H) were simulated using k⁺_poly = 1/s and k⁻_poly = 0.1/s. The visitation analysis (A, E), average rates (C, G) extracted from the average traces (B, F) and dwell-time analysis (D, H) were applied to data with 100% labeling efficiency and no photobleaching (blue data). The average trace of restricted growth reaches only 2.5 monomers, despite possible growth until 4 monomers, because of the equilibrium between the on- and the off-rate. The effect of a photobleaching rate of 0.01/s does not significantly influence the extracted rates and the visitation analysis (red data). The combination of stochastic labeling of 30% and photobleaching with a rate of 0.01/s affects all analysis methods (yellow). Error bars for the dwell-time analysis represent the 95% confidence intervals of the exponential fit.