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Enhanced multiplexing in mass cytometry using osmium and ruthenium tetroxide species
Author(s) -
Catena Raúl,
Özcan Alaz,
Zivanovic Nevena,
Bodenmiller Bernd
Publication year - 2016
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.22848
Subject(s) - osmium tetroxide , ruthenium , mass cytometry , osmium , flow cytometry , multiplexing , chemistry , cytometry , computer science , biology , microbiology and biotechnology , biochemistry , physics , telecommunications , catalysis , optics , electron microscope , gene , phenotype
Mass cytometry facilitates high‐dimensional, quantitative, single‐cell analysis. The method for sample multiplexing in mass cytometry, called mass‐tag cellular barcoding (MCB), relies on the covalent reaction of bifunctional metal chelators with intracellular proteins. Here, we describe the use of osmium and ruthenium tetroxides (OsO 4 and RuO 4 ) that bind covalently with fatty acids in the cellular membranes and aromatic amino acids in proteins. Both OsO 4 and RuO 4 rapidly reacted and allowed for MCB with live cells, crosslinked cells, and permeabilized cells. Given the covalent nature of the labeling reaction, isotope leaching was not observed. OsO 4 and RuO 4 were used in a 20‐sample barcoding protocol together with palladium isotopes. As mass channels occupied by osmium and ruthenium are not used for antibody detection the number of masses effectively utilized in a single experiment is expanded. OsO 4 and RuO 4 can therefore be used as MCB reagents for a wide range of mass cytometry workflows. © 2016 International Society for Advancement of Cytometry