Titration of Mass Cytometry Reagents

Caryn van Vreden; Paula Niewold; Helen M McGuire; Barbara Fazekas de St Groth

doi:10.1007/978-1-4939-9454-0_6

Titration of Mass Cytometry Reagents

Methods Mol Biol. 2019:1989:83-92. doi: 10.1007/978-1-4939-9454-0_6.

Authors

Caryn van Vreden^{1

2

3}, Paula Niewold^{3

4}, Helen M McGuire^{5

6

7}, Barbara Fazekas de St Groth^{8

9

10}

Affiliations

¹ Sydney Cytometry, The University of Sydney and Centenary Institute, Sydney, Australia.
² Ramaciotti Facility for Human Systems Biology, The University of Sydney, Sydney, Australia.
³ Charles Perkins Centre, The University of Sydney, Sydney, Australia.
⁴ Discipline of Pathology, School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia.
⁵ Ramaciotti Facility for Human Systems Biology, The University of Sydney, Sydney, NSW, Australia.
⁶ Charles Perkins Centre, The University of Sydney, Sydney, NSW, Australia.
⁷ Discipline of Pathology, The University of Sydney, Camperdown, NSW, Australia.
⁸ Ramaciotti Facility for Human Systems Biology, The University of Sydney, Sydney, Australia. [email protected].
⁹ Charles Perkins Centre, The University of Sydney, Sydney, Australia. [email protected].
¹⁰ Discipline of Pathology, School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia. [email protected].

PMID: 31077100
DOI: 10.1007/978-1-4939-9454-0_6

Abstract

Mass cytometry (MC) is a powerful research tool enabling high-dimensional analysis of single cells in suspension and within tissue sections following laser ablation. Here we describe the procedure of titrating metal-conjugated antibodies, to ensure that optimal levels of staining are achieved while minimizing nonspecific signals that may occur at high concentrations.

Keywords: Antibody titration; Best practices; Conjugation; CyTOF; Helios; Mass cytometry.

MeSH terms

Antibodies, Monoclonal
Cells / cytology*
Chelating Agents / chemistry
Flow Cytometry / methods*
Humans
Immunoconjugates / chemistry
Immunoglobulins / immunology
Immunophenotyping / methods*
Indicators and Reagents
Lasers
Mass Spectrometry / methods*
Metals / chemistry
Single-Cell Analysis / methods*

Substances

Antibodies, Monoclonal
Chelating Agents
Immunoconjugates
Immunoglobulins
Indicators and Reagents
Metals