Reducing Bias in Mass Cytometry Data through Corrections for Spillover and Nonspecific Binding.

ABSTRACT: Mass cytometry can facilitate the study of over 40 protein markers in single cells, making it an ideal technique to investigate complex biological systems. The use of antibodies tagged with metal isotopes as probes makes mass cytometry measurements susceptible to isotopic spillover. In this report, we introduce ccSpill, an instrument-specific matrix built from calibration curves of metal-labeled antibodies and solutions of pure metals that can be applied to single-cell data to remove spillover. ccSpill is unique to each lab, and each set of reporter antibodies and is compatible with isotype-based nonspecific binding (NSB) corrections. We demonstrate that combining ccSpill with NSB corrections is integral to the optimization of sample preparation procedures that involve challenging antibodi

SUBMITTER: Elbein K 

PROVIDER: S-EPMC12758627 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature