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High-Throughput Mass Cytometry Staining for Immunophenotyping Clinical Samples

Emily M. Thrash, Katja Kleinsteuber, Emma Hathaway, Matthew Nazzaro, Eric Haas, F. Stephen Hodi, Mariano Severgnini

2020STAR Protocols34 citationsDOIOpen Access PDF

Abstract

As mass cytometry (MC) is implemented in clinical settings, the need for robust, validated protocols that reduce batch effects between samples becomes increasingly important. Here, we present a streamlined MC workflow for high-throughput staining that generates reproducible data for up to 80 samples in a single experiment by combining reference sample spike-in and palladium-based mass-tag cell barcoding. Although labor intensive, this workflow decreases experimental variables and thus reduces technical error and mitigates batch effects.

Topics & Concepts

Mass cytometryWorkflowThroughputImmunophenotypingComputer scienceStainingSample (material)CytometryFlow cytometryChromatographyBiologyChemistryDatabaseMedicineMolecular biologyPathologyTelecommunicationsWirelessPhenotypeGeneBiochemistrySingle-cell and spatial transcriptomicsCell Image Analysis TechniquesAdvanced Fluorescence Microscopy Techniques
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