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. 2019 Aug 23;9:12297. doi: 10.1038/s41598-019-48814-w

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© The Author(s) 2019

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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Automated real-time imaging of human induced pluripotent stem cell suspension cultures. (A) Outline of the cell suspension circulation. Cell suspension was sampled from the reactor by peristaltic pumping and led through a flow chamber above the microscope lens. Images were acquired via CCD camera while the suspension is in flow. A computer was used for at-line image processing, impeller speed control, and pH and oxygen measurement recording. (B) Suspension culture samples were investigated for lactate dehydroxynase activity and (C) cell viability to convey potential cell damage caused by the peristaltic pump during circulation. (D) Processing of suspension culture images provided aggregate size distributions. Here, the mean aggregate diameter µ and its standard deviation σ are used as numerical readout parameters. Automated real-time imaging allowed for detailed characterization of the aggregate formation during bioreactor culture by plotting (E) the average aggregate diameter µ and (F) the standard deviation σ of the aggregate diameter µ as a function of cultivation time. Error bars in all subfigures indicate mean ± SD from n = 3 experiments.