title: Cell size assays for mass cytometry.
abstract: Mass cytometry offers the advantage of allowing the simultaneous measurement of a greater number parameters than conventional flow cytometry. However, to date, mass cytometry has lacked a reliable alternative to the light scatter properties that are commonly used as a cell size metric in flow cytometry (forward scatter intensity-FSC). Here, we report the development of two plasma membrane staining assays to evaluate mammalian cell size in mass cytometry experiments. One is based on wheat germ agglutinin (WGA) staining and the other on Osmium tetroxide (OsO4) staining, both of which have preferential affinity for cell membranes. We first perform imaging and flow cytometry experiments to establish a relationship between WGA staining intensity and traditional measures of cell size. We then incorporate WGA staining in mass cytometry analysis of human whole blood and show that WGA staining intensity has reproducible patterns within and across immune cell subsets that have distinct cell sizes. Lastly, we stain PBMCs or dissociated lung tissue with both WGA and OsO4; mass cytometry analysis demonstrates that the two staining intensities correlate well with one another. We conclude that both WGA and OsO4may be used to acquire cell size-related parameters in mass cytometry experiments, and expect these stains to be broadly useful in expanding the range of parameters that can be measured in mass cytometry experiments. © 2016 International Society for Advancement of Cytometry.
authors: Department of Pharmacology and Systems Therapeutics, Icahn School of Medicine at Mount Sinai, New York, New York, 10029.
authors: Alan D
authors: AD
authors: Stern
authors: Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, 10029.|Icahn School of Medicine at Mount Sinai, Human Immune Monitoring Core, New York, New York, 10029.
authors: Adeeb H
authors: AH
authors: Rahman
authors: Department of Pharmacology and Systems Therapeutics, Icahn School of Medicine at Mount Sinai, New York, New York, 10029.
authors: Marc R
authors: MR
authors: Birtwistle
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000648:ultrastructure*; D048429:Cell Size; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D009993:Osmium Tetroxide / Q000737:chemistry; D014909:Wheat Germ Agglutinins / Q000737:chemistry
chemical_list: D014909:Wheat Germ Agglutinins; D009993:Osmium Tetroxide
keywords: CyTOF; HEK293; U87; WGA; cell size; flow cytometry; mass cytometry; whole blood