Simplified mass cytometry protocol for in-plate staining, barcoding, and cryopreservation of human PBMC samples in clinical trials

Charlotte Abrecht; Margaret Hallisey; Jenna Dennis; Matthew Nazzaro; Martha Brainard; Emma Hathaway; Abigail N. Schork; F. Stephen Hodi; Mariano Severgnini; Joanna Baginska

STAR Protocols (Jun 2022)

Simplified mass cytometry protocol for in-plate staining, barcoding, and cryopreservation of human PBMC samples in clinical trials

Charlotte Abrecht,
Margaret Hallisey,
Jenna Dennis,
Matthew Nazzaro,
Martha Brainard,
Emma Hathaway,
Abigail N. Schork,
F. Stephen Hodi,
Mariano Severgnini,
Joanna Baginska

Affiliations

Charlotte Abrecht: Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, MA 02215, USA
Margaret Hallisey: Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, MA 02215, USA; Corresponding author
Jenna Dennis: Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, MA 02215, USA
Matthew Nazzaro: Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, MA 02215, USA
Martha Brainard: Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, MA 02215, USA
Emma Hathaway: Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, MA 02215, USA
Abigail N. Schork: Longwood Medical Area CyTOF Core, Dana-Farber Cancer Institute, Boston, MA 02215, USA
F. Stephen Hodi: Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, MA 02215, USA
Mariano Severgnini: Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, MA 02215, USA
Joanna Baginska: Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, MA 02215, USA; Corresponding author

Journal volume & issue: Vol. 3, no. 2
p. 101362

Abstract

Read online

Summary: With the increasing use of mass cytometry in clinical research, a simplified and standardized protocol for immunophenotyping human peripheral blood mononuclear cells (PBMCs) in clinical trials is needed. We present a simplified in-plate staining protocol for up to 80 samples, for laboratories of all mass cytometry expertise levels, aimed to generate reproducible datasets for large clinical cohorts. In this protocol, we provide details on the requirements to obtain meaningful results, spanning from sample quality, barcoding, and batch-freezing of stained samples.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

About the journal

Abstract

Keywords