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. 2020 Jan 24;10:1149. doi: 10.1038/s41598-019-57387-7

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© The Author(s) 2020

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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Flow cytometry based autofluorescent (AF) RSCs sorting. Here, FCS Express 6.06.0014 (De Novo Software) generated two-parameter density plot shows the density of cells at a given position represented by different colours. (A) Sampling of coelomocytes and RSCs enrichment. The density plot represents; (B) Forward/Scatter profile of live unstained coelomocyte. (C) Microscopic image of RSCs and other cells before sorting. (D) Gating of singlets allowing discrimination of doublets and (E) Gating of distinct fluorescent RSCs population represented by the red box. (F) Histogram of fluorescent RSCs population. The histogram marker presents the selective population to be sorted. (G) Microscopic image of pure AF-RSCs after sorting. Microscopic image scale bar 50 µm.