Osmium-Labeled Microspheres for Bead-Based Assays in Mass Cytometry

Polystyrene beads are broadly applied in flow cytometry. Implementing bead-based assays in mass cytometry is desired but hampered by the lack of an elemental label required for their detection. In this study, we introduce stable osmium tetroxide labeling as a universal approach for generating functi...

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Bibliographic Details
Published in:The Journal of immunology (1950) 2019-05, Vol.202 (10), p.3103-3112
Main Authors: Budzinski, Lisa, Schulz, Axel R, Baumgart, Sabine, Burns, Tyler, Rose, Thomas, Hirseland, Heike, Mei, Henrik E
Format: Article
Language:English
Subjects:
Adult
Aged
Female
Flow Cytometry
Gene Expression Regulation - immunology
HLA-DR Antigens - blood
HLA-DR Antigens - immunology
Humans
Immunologic Memory
Lupus Erythematosus, Systemic - blood
Lupus Erythematosus, Systemic - immunology
Lupus Erythematosus, Systemic - pathology
Male
Microspheres
Middle Aged
Osmium - chemistry
Polystyrenes - chemistry
Staining and Labeling
T-Lymphocytes, Regulatory - immunology
T-Lymphocytes, Regulatory - metabolism
T-Lymphocytes, Regulatory - pathology
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Summary:Polystyrene beads are broadly applied in flow cytometry. Implementing bead-based assays in mass cytometry is desired but hampered by the lack of an elemental label required for their detection. In this study, we introduce stable osmium tetroxide labeling as a universal approach for generating functionalized beads readily detectable by mass cytometry. We demonstrate the utility of osmium-labeled beads for signal spillover compensation in mass cytometry, and, strikingly, their application in quantitative Ab-binding capacity assays combined with high-dimensional profiling of human PBMC enabled the systematic assessment of receptor expression profiles across large numbers of cellular phenotypes. This analysis confirmed increased monocytic Siglec-1 expression in active systemic lupus erythematosus patients and, additionally, revealed interrelated reductions of CD4 expression by regulatory and memory CD4 T cells and HLA-DR expression by myeloid dendritic cells, pointing toward defective cross-talk at the immunological synapse that may limit immune responses in systemic lupus erythematosus. By converting conventional flow cytometry beads into beads suitable for mass cytometry, our approach paves the way toward the broad implementation of bead-based assays in high-dimensional cell profiling studies by mass cytometry in biomedical research.
ISSN:0022-1767
1550-6606
DOI:10.4049/jimmunol.1801640