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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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Experimental data of 30% stochastically labeled actin-Cy5 growing on Cappuccino and Spire-ABCD. A comparison between a visitation analysis, dwell-time analysis and average rate analysis is performed for the measurements with Cappuccino (A–D, green) and Spire-ABCD (E–G, blue). (A, E): The visitation analysis, (B, F) average intensity, (C, G) fractional average-rate analysis and (D, H) dwell-time analysis showing the first 5 monomer association steps from 700 individual traces are plotted. The fractional average rates have been calculated using the time until the average trace reached the intensity change corresponding to 0.1 monomers (i.e. between 0 and 0.1, between 0.1 and 0.2, between 0.2 and 0.3, etc.). Fractional average rates are 10 times faster than average rates because the average intensity of only 1/10 of a monomer has to be reached. The experimentally determined photobleaching rate was 0.025/s.