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. 2022 Aug 15;12:13818. doi: 10.1038/s41598-022-17548-7

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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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Evaluation of contractile force from wrinkles formed on the substrate. (a) How wrinkles on the substrate are extracted. The source movie was divided into frames. The wrinkles were extracted from the difference between images of contracted myotubes and relaxed myotubes. The difference image is binarized so the wrinkles appear as brighter lines. (b) Extracted wrinkles superimposed on the source movie (see Supplementary Movie 3), shown as white lines. (c) The length of wrinkles was found in pixels using ImageJ-Fiji. The total length of wrinkles generated by the first three contractions of myotubes was defined as a “force index”.