Mass Cytometry Analysis of Whole Blood Response to an Allergen

Daniel Geanon, Charuta Agashe, Adeeb Rahman, M. Cecilia Berin*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Mass cytometry allows for the use of highly multiplexed antibody panels due to the lack of spill-over between channels detected by mass spectrometry. An advantage over fluorescence cytometry is the relative lack of background, which provides excellent resolution for detection of phosphoproteins and quantification of cell signaling. We have applied mass cytometry to the analysis of whole blood staining after ex vivo stimulation with peanut allergen (Tordesillas et al., J Allergy Clin Immunol 138:1741–4.e9, 2016). This allows for high-dimensional analysis of basophil activation, and analysis of the entire composition of the blood compartment in response to allergen exposure. Here, we describe our optimized protocol for activation and staining of whole blood for mass cytometry analysis that is currently in use in multicenter clinical trials. The protocol can be easily adopted to analyze blood leukocytes in other diseases, including asthma.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages269-280
Number of pages12
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2506
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Basophil
  • BAT
  • Peanut allergy

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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