title: A platinum-based covalent viability reagent for single-cell mass cytometry.
abstract: In fluorescence-based flow cytometry, cellular viability is determined with membrane-impermeable fluorescent reagents that specifically enter and label plasma membrane-compromised nonviable cells. A recent technological advance in flow cytometry uses antibodies conjugated to elemental metal isotopes, rather than to fluorophores, to allow signal detection by atomic mass spectrometry. Unhampered by the limitations of overlapping emission fluorescence, mass cytometry increases the number of parameters that can be measured in single cells. However, mass cytometry is unable to take advantage of current fluorescent viability dyes. An alternative methodology was therefore developed here in which the platinum-containing chemotherapy drug cisplatin was used to resolve live and dead cells by mass cytometry. In a 1-min incubation step, cisplatin preferentially labeled nonviable cells from both adherent and suspension cultures, resulting in a platinum signal quantifiable by mass cytometry. This protocol was compatible with established sample processing steps for intracellular cytometry. Furthermore, the live/dead ratios were comparable between mass- and fluorescence-based cytometry. Importantly, although cisplatin is a known DNA-damaging agent, a 1-min "pulse" of cisplatin did not induce observable DNA damage or apoptotic responses even within 6-h post-exposure. Cisplatin can therefore be used as a viability reagent for a wide range of mass cytometry protocols.
authors.affiliation: Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Harris G
authors.initials: HG
authors.lastname: Fienberg
authors.forename: Erin F
authors.initials: EF
authors.lastname: Simonds
authors.forename: Wendy J
authors.initials: WJ
authors.lastname: Fantl
authors.forename: Garry P
authors.initials: GP
authors.lastname: Nolan
authors.forename: Bernd
authors.initials: B
authors.lastname: Bodenmiller
mesh_terms: D002448:Cell Adhesion; D002463:Cell Membrane Permeability; D002470:Cell Survival; D002945:Cisplatin / Q000737:chemistry*; D004247:DNA / Q000032:analysis / Q000737:chemistry; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D010984:Platinum / Q000737:chemistry*; D059010:Single-Cell Analysis; D013194:Staining and Labeling / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D005456:Fluorescent Dyes; D010984:Platinum; D004247:DNA; D002945:Cisplatin