Chevrier S Crowell HL Zanotelli VRT Engler S Robinson MD Bodenmiller B 2018 The advent of mass cytometry increased the number of parameters measured at the single-cell level while decreasing the extent of crosstalk between channels relative to dye-based flow cytometry. Although reduced, spillover still exists in mass cytometry data, and minimizing its effect requires considerable expert knowledge and substantial experimental effort. Here, we describe a novel bead-based compensation workflow and R-based software that estimates and corrects for interference between channels. We performed an in-depth characterization of the spillover properties in mass cytometry, including limitations defined by the linear range of the mass cytometer and the reproducibility of the spillover over time and across machines. We demonstrated the utility of our method in suspension and imaging mass cytometry. To conclude, our approach greatly simplifies the development of new antibody panels, increases flexibility for antibody-metal pairing, opens the way to using less pure isotopes, and improves overall data quality, thereby reducing the risk of reporting cell phenotype artifacts. application/pdf https://sonar.ch/global/documents/99094 eng info:eu-repo/semantics/altIdentifier/doi/10.1016/j.cels.2018.02.010 info:eu-repo/semantics/altIdentifier/pmid/29605184 info:eu-repo/semantics/openAccess Cell systems. - 2018 CyTOF channel interference compensation imaging mass cytometry mass cytometry signal crosstalk spillover Antibodies Breast Neoplasms Female Flow Cytometry Humans Image Cytometry Immunophenotyping Reproducibility of Results Signal-To-Noise Ratio Single-Cell Analysis Software Suspensions Compensation of Signal Spillover in Suspension and Imaging Mass Cytometry. http://purl.org/coar/resource_type/c_6501