Mass Cytometry for the Characterization of Individual Cell Types in Ovarian Solid Tumors

Abstract

Mass cytometry aka Cytometry by Time-Of-Flight (CyTOF) is one of several recently developed multiparametric single-cell technologies designed to address cellular heterogeneity within healthy and diseased tissue. Mass cytometry is an adaptation of flow cytometry in which antibodies are labeled with stable heavy metal isotopes and the readout is by time-of-flight mass spectrometry. With minimal spillover between channels, mass cytometry enables readouts of up to 60 parameters per single cell. Critically, mass cytometry can identify minority cell populations that are lost in bulk tissue analysis. Mass cytometry has been used to great effect for the study of immune cells. We have extended its use to examine single cells within disaggregated solid tissues, specifically freshly resected tubo-ovarian high-grade serous tumors. Here we detail our protocols designed to ensure the production of high-quality single-cell datasets. The methodology can be modified to accommodate the study of other solid tissues.

Keywords: Antibody conjugation; Cisplatin; CyTOF; Heavy metal isotope; High-grade serous ovarian cancer; Lanthanides; Live–dead stain; Mass cytometry; Maxpar labeling kits; Palladium barcoding; Rhodium; Single-cell dissociation; Solid tumors.