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. 2020 Jul 20;10:11991. doi: 10.1038/s41598-020-68770-0

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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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Process and Rationale Illustration. (A) Rats were injured using the controlled cortical impact method, to create a precise injury at one hemisphere. (B) Each hemisphere was processed to a separate sample, by mechanically and enzymatically digesting the tissue into single cell suspension (D), myelin removal (E) and microglial (CD11bc) enrichment (F), which is the final sample that was later analyzed with flow cytometry. (G) Considering the morphological changes and the changes in markers expression we used a validated multi-parametric flow cytometry panel to evaluate all parameters together to create a profile of the activated microglia at 24 after injury. Made in ©BioRender—biorender.com.