Binek, AleksandraBarbas Arribas, CoralRupérez Pascualena, Francisco JavierGodzien, Joanna BarbaraNúñez, VanessaJorge, InmaculadaRicote, MercedesVázquez, JesúsRojo Blanco, DavidUniversidad San Pablo-CEU. Facultad de Farmacia. Departamento de Química y Bioquímica2024-01-182024-01-182019-01-04Aleksandra Binek, David Rojo, Joanna Godzien, Francisco Javier Rupérez, Vanessa Nuñez, Inmaculada Jorge, Mercedes Ricote, Jesús Vázquez, and Coral Barbas. Flow Cytometry Has a Significant Impact on the Cellular Metabolome. Journal of Proteome Research 2019 18 (1), 169-181. DOI: 10.1021/acs.jproteome.8b004721535-3907http://hdl.handle.net/10637/14995This is the peer reviewed version of the article which has been published in final form at: https://doi.org/10.1021/acs.jproteome.8b00472The characterization of specialized cell subpopulations in a heterogeneous tissue is essential for understanding organ function in health and disease. A popular method of cell isolation is fluorescence-activated cell sorting (FACS) based on probes that bind surface or intracellular markers. In this study, we analyze the impact of FACS on the cell metabolome of mouse peritoneal macrophages. Compared with directly pelleted macrophages, FACS-treated cells had an altered content of metabolites related to the plasma membrane, activating a mechanosensory signaling cascade causing inflammation-like stress. The procedure also triggered alterations related to energy consumption and cell damage. The observed changes mostly derive from the physical impact on cells during their passage through the instrument. These findings provide evidence of FACS-induced biochemical changes, which should be taken into account in the design of robust metabolic assays of cells separated by flow cytometry.esopen accessSortingSorted cellsMetabolome profileFluorescence activated cell sorting (FACS)LC39 MSMetabolomicsMultiplatform analysisArtículo10.1021/acs.jptoteome.8b00472https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es