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. 2021 Jan 18;11:1662. doi: 10.1038/s41598-021-81217-4

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© The Author(s) 2021

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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R-integrated pixel intensity (PIXI) analysis from brightfield images as a reproducible image analysis tool. (a) Curve fit of the number of plated cells (50–300 × 10³ cells/well, n = 14–16) versus total brightfield pixel intensity values, indicated by Pearson correlation coefficients (R²). Data is represented as mean ± s.d.. Calibration curves were used to transform total pixel intensity values back into cell numbers using second-order polynomial regression analysis. (b) Coefficients of variation (CV in %) within each donor for the cell counts of the total PBMC pool before seeding (n = 8, represented as mean) and the brightfield image PIXI analysis (represented as median ± range; means are visualized by χ-bar).