High-parameter immune profiling is a cornerstone in translational and clinical research used to quantify changes in diverse immune cell populations and states over time. CyTOF® mass cytometry is a highly multiplexed, single-cell analysis platform that uses isotopically pure metal-labeled antibodies. A significant advantage of CyTOF is its ability to resolve more than 40 markers in a single panel without the need for compensation, making mass cytometry an ideal solution for routine enumeration and phenotyping of immune cells. The Maxpar® Direct™ Immune Profiling Assay™ and Maxpar Pathsetter™ software were developed as a sample-to-answer solution for human immune profiling using mass cytometry. The Maxpar Direct Immune Profiling Assay kit includes an optimized 30-marker panel contained in a lyophilized single-tube format. Maxpar Pathsetter is an automated software to report population statistics, stain assessments, and relevant data plots. The Maxpar Direct Immune Profiling System was originally validated for Helios™ mass cytometers, but now acquisition can be simplified with an automated acquisition system for mass cytometers.
Using the automated acquisition system to acquire samples stained with the Maxpar Direct Immune Profiling Assay followed by analysis in Maxpar Pathsetter provides a streamlined workflow for immune profiling. Here, we provide performance assessment of this automated acquisition system compared to the single sample acquisition system. Human whole blood and PBMC samples were stained with the Maxpar Direct Immune Profiling Assay. Normalized files were then analyzed using Maxpar Pathsetter. We show a high degree of repeatability of this assay when acquired using both acquisition systems. Deming regression was performed to demonstrate improved staining assessment and comparable population frequencies. Overall, combining the automated acquisition system and the Maxpar Direct Immune Profiling System enables researchers to simplify and streamline high-parameter immunophenotyping of human whole blood and PBMC samples while accurately and reproducibly monitoring changes in immune cell subsets.