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. 2021 Dec 14;11:23935. doi: 10.1038/s41598-021-03442-1

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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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Different room temperature pre-incubation periods affect cell plating homogeneity across the well in 96-well plates. (a) Full well scan acquired using the full-well scan function on the EVOS FL Auto microscope with a ×10 objective lens. Yellow box indicates the region where cells are sparsely distributed. (b) Schematic illustration of well divisions into five regions for analysis. (c) Cell plating homogeneity was determined as the standard deviation between each region. This was used to evaluate the efficacy of pre-incubation for different time periods. Each bar represents the mean (n = 3) ± SD. (d) Cell plating homogeneity defined as the standard deviation between each region was used to evaluate the efficacy of pre-incubation for different time periods. Each bar represents the mean (n = 5) ± SD. A one-way ANOVA test and Tukey’s post-hoc analysis was performed to evaluate statistical significance where **p < 0.01 and *p < 0.05.