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Updated: Sep 6, 2025
Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
Published on: June 26, 2018
Mass Cytometry Analysis of Whole Blood Response to an Allergen
Daniel Geanon1, Charuta Agashe1, Adeeb Rahman1
1Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Insights
Mass cytometry offers high-dimensional analysis of immune cells, like basophils, following allergen exposure. This optimized protocol enhances whole blood analysis for clinical trials and other diseases such as asthma.
Area of Science:
- Immunology
- Biotechnology
- Analytical Chemistry
Background:
- Mass cytometry provides highly multiplexed antibody panels with minimal signal spill-over.
- It offers superior resolution for phosphoprotein detection and cell signaling quantification compared to fluorescence cytometry.
- Previous work applied mass cytometry to analyze basophil activation after peanut allergen stimulation.
Purpose of the Study:
- To describe an optimized protocol for whole blood activation and staining for mass cytometry.
- To enable high-dimensional analysis of immune cell responses to allergen exposure.
- To facilitate the application of mass cytometry in multicenter clinical trials and other disease analyses.
Main Methods:
- Whole blood samples were stimulated ex vivo with peanut allergen.
- Optimized staining and activation protocols were developed for mass cytometry.
- Mass cytometry was used for high-dimensional analysis of blood cell composition and basophil activation.
Main Results:
- The protocol allows for detailed analysis of basophil activation.
- It enables comprehensive assessment of the entire blood compartment composition.
- The method provides high-dimensional data for immune response characterization.
Conclusions:
- An optimized mass cytometry protocol for whole blood analysis was established.
- This protocol is suitable for use in multicenter clinical trials.
- The method can be adapted for studying blood leukocytes in various diseases, including asthma.
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.
