article published yes Nicole Amberg author 4CD6AAC6-F248-11E8-B48F-1D18A9856A870000-0002-3183-8207 Giselle T Cheung author 471195F6-F248-11E8-B48F-1D18A9856A870000-0001-8457-2572 Simon Hippenmeyer author 37B36620-F248-11E8-B48F-1D18A9856A870000-0003-2279-1061 SiHi department Role of Eed in neural stem cell lineage progression project ISTplus - Postdoctoral Fellowships project Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular Mechanisms of Neural Stem Cell Lineage Progression project Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development project Mosaic analysis with double markers (MADM) technology enables the generation of genetic mosaic tissue in mice and high-resolution phenotyping at the individual cell level. Here, we present a protocol for isolating MADM-labeled cells with high yield for downstream molecular analyses using fluorescence-activated cell sorting (FACS). We describe steps for generating MADM-labeled mice, perfusion, single-cell suspension, and debris removal. We then detail procedures for cell sorting by FACS and downstream analysis. This protocol is suitable for embryonic to adult mice. For complete details on the use and execution of this protocol, please refer to Contreras et al. (2021).1 https://research-explorer.ista.ac.at/download/14683/17260/2024_STARProtoc_Amberg.pdf application/pdfno Elsevier2024 eng General Immunology and MicrobiologyGeneral BiochemistryGenetics and Molecular BiologyGeneral Neuroscience 2666-1667 3807013710.1016/j.xpro.2023.102771 51 Amberg, Nicole, Giselle T Cheung, and Simon Hippenmeyer. “Protocol for Sorting Cells from Mouse Brains Labeled with Mosaic Analysis with Double Markers by Flow Cytometry.” <i>STAR Protocols</i>. Elsevier, 2024. <a href="https://doi.org/10.1016/j.xpro.2023.102771">https://doi.org/10.1016/j.xpro.2023.102771</a>. Amberg, Nicole, et al. “Protocol for Sorting Cells from Mouse Brains Labeled with Mosaic Analysis with Double Markers by Flow Cytometry.” <i>STAR Protocols</i>, vol. 5, no. 1, 102771, Elsevier, 2024, doi:<a href="https://doi.org/10.1016/j.xpro.2023.102771">10.1016/j.xpro.2023.102771</a>. N. Amberg, G. T. Cheung, and S. Hippenmeyer, “Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry,” <i>STAR Protocols</i>, vol. 5, no. 1. Elsevier, 2024. Amberg N, Cheung GT, Hippenmeyer S. 2024. Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry. STAR Protocols. 5(1), 102771. N. Amberg, G.T. Cheung, S. Hippenmeyer, STAR Protocols 5 (2024). Amberg N, Cheung GT, Hippenmeyer S. Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry. <i>STAR Protocols</i>. 2024;5(1). doi:<a href="https://doi.org/10.1016/j.xpro.2023.102771">10.1016/j.xpro.2023.102771</a> Amberg, N., Cheung, G. T., &#38; Hippenmeyer, S. (2024). Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry. <i>STAR Protocols</i>. Elsevier. <a href="https://doi.org/10.1016/j.xpro.2023.102771">https://doi.org/10.1016/j.xpro.2023.102771</a> 146832023-12-13T11:48:05Z2025-04-15T08:23:06Z