An automated approach to high-plex cytometric immunophenotyping with CyTOF XT

Abstract

Abstract CyTOF® mass cytometry is a single-cell analysis platform that uses isotope-tagged antibodies to resolve 50-plus markers in a single tube without signal compensation, making CyTOF ideal for routine immunophenotyping. CyTOF XT™, the latest CyTOF system, features automated sample acquisition. Stained samples were acquired in parallel using the automated CyTOF XT system and manually, using the Helios™ system, to assess performance of the automated system. Multiple suspension mass cytometry staining workflows were evaluated. Population frequencies and resolution indices for markers were assessed by manual gating. There was no significant difference between population frequencies analyzed between the two CyTOF systems. On average, samples acquired on CyTOF XT resulted in greater resolution between positive and negative populations compared to Helios. The Maxpar® Direct™ Immune Profiling System, which comprises the Maxpar® Direct™ Immune Profiling Assay™ and Maxpar Pathsetter™ software, was also compared on the CyTOF XT and Helios systems. The Maxpar Direct Immune Profiling Assay includes a 30-marker panel in a dry, single-tube format for staining human whole blood or PBMC. Maxpar Pathsetter automates reporting of population statistics and stain assessments for the panel. Maxpar Pathsetter showed comparable population frequencies between the two CyTOF systems and improved staining assessment on CyTOF XT. Overall, these studies find that the CyTOF XT system generates better signal resolution than the Helios system. Automated acquisition by CyTOF XT enables researchers to accurately and reproducibly streamline human immunophenotyping. For Research Use Only. Not for use in diagnostic procedures.