Alternative methods of viability determination in single cell mass cytometry

The identification and discrimination of viable cells is important to understand how experimental variables may influence biochemical processes such as cell metabolism, cell cycle, and signaling pathways. Cisplatin is commonly used as a viability stain in mass cytometry studies, however, recent work...

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Bibliographic Details
Published in:Cytometry. Part A 2021-10, Vol.99 (10), p.1042-1053
Main Authors: Devine, Raymond D., Alkhalaileh, Hussam S., Lyberger, Justin M., Behbehani, Gregory K.
Format: Article
Language:English
Subjects:
Antibodies
Antigens
Cell cycle
cell death
cell viability
chemotherapy
Chlorides
Cisplatin
Cytometry
Dimensional analysis
Fractions
Glyceraldehyde-3-phosphate dehydrogenase
Hafnium
Histone H1
Isotopes
Markers
mass cytometry
Metabolism
Niobium
Platinum
Platinum isotopes
Populations
Proteins
Salts
Staining
Stains
Stains & staining
Viability
Zirconium
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Summary:The identification and discrimination of viable cells is important to understand how experimental variables may influence biochemical processes such as cell metabolism, cell cycle, and signaling pathways. Cisplatin is commonly used as a viability stain in mass cytometry studies, however, recent work by Mei et al. has demonstrated that cisplatin can also be used to label antibodies, complicating the simultaneous use of the platinum measurement channels for both antibody and viability staining. This study demonstrates that other metal salts (hafnium chloride, niobium chloride, and zirconium chloride) can serve as substitutes for cisplatin in viability staining. These stains yield similar fractions of live and dead cells and stain the same dead cells in parallel high parameter analyses. In addition, this study demonstrates how a variety of protein antigen viability markers (pRb, Ki‐67, Histone H1, cleaved PARP, and GAPDH) can be used to discriminate live and dead cell populations, without the need for a separate viability staining step. As few as two of these protein antigen viability markers can help identify live and dead cell populations in fixed samples and can identify the same viable cells in high dimensional analyses with or without use of viability stain information. This study demonstrates several alternative approaches to mass cytometry viability assessment that can facilitate use of platinum isotopes for antibody staining and enables identification of live and dead cell populations in samples for which a separate viability stain is not practical.
ISSN:1552-4922
1552-4930
DOI:10.1002/cyto.a.24308