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URL: https://dx.doi.org/10.17504/protocols.io.78vhrw6

Authors: Konstantin J. Bode, Stefanie Mueller, Matthias Schweinlin, Marco Metzger, Thomas Brunner

Group: BioTechniques

Summary: This protocol describes a novel fluorometric method for the quantitative measurement of specific organoid cell death. Organoids are stained simultaneously with the cell impermeable nuclear dye propidium iodide and cell permeable Hoechst 33342. While Hoechst allows in-well normalization to cell numbers, propidium iodide detects relative proportion of dead cells independent of hydrogel. Measurement and analysis time, as well as usability are drastically improved in comparison to other established methods. Parallel multiplexing of this method with established assays measuring mitochondrial activity further enhances its applicability in personalized medicine and drug discovery.

Affiliations: Chair of Biochemical Pharmacology, Department of Biology, University of Konstanz, Konstanz, Germany; Co-operative research training school 'Advanced in-vitro test systems for the analysis of cell-chemical interactions in drug discovery and environmental safety (InViTe)', University of Konstanz, Konstanz, Germany, Chair of Biochemical Pharmacology, Department of Biology, University of Konstanz, Konstanz, Germany, Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Würzburg, Würzburg, Germany, Translational Centre Regenerative Medicine TLC-RT, Fraunhofer Institute for Silicate Research (ISC), Würzburg, Germany, Chair of Biochemical Pharmacology, Department of Biology, University of Konstanz, Konstanz, Germany

External URL: https://www.future-science.com/doi/10.2144/btn-2019-0023

Version: 1

Publication Date: 2019

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