High-Parameter Immune Profiling with CyTOF.
Mass cytometry, or CyTOF, is a useful technology for high-parameter single-cell phenotyping, especially from suspension cells such as blood or PBMC. It is particularly appealing to monitor the systemic immune changes that could accompany cancer immunotherapy. Here we present a reference panel for identification of all major immune cell populations, with flexibility for addition of trial-specific markers. We also describe best-practice measures for minimizing and tracking batch variability. These include: sample barcoding, use of spiked-in reference cells, and lyophilization of the antibody cocktail. Our protocol assumes the use of cryopreserved PBMC, both for convenience of batching samples and for maximum comparability across patients and time points. Finally, we show an option for automated analysis using the Astrolabe platform (Astrolabe Diagnostics, Inc.).
['Sahaf|Bita|B|', 'Rahman|Adeeb|A|', 'Maecker|Holden T|HT|', 'Bendall|Sean C|SC|']
D000906:Antibodies D001800:Blood Specimen Collection D002469:Cell Separation D005434:Flow Cytometry D005612:Freeze Drying D017408:Guidelines as Topic D006801:Humans D007963:Leukocytes, Mononuclear D013058:Mass Spectrometry D009369:Neoplasms D040901:Proteomics D059010:Single-Cell Analysis
Mass cytometry Differential analysis Multiplexed Phenotypic markers Subpopulation