Mass Cytometry Analysis of Whole Blood Response to an Allergen

Daniel Geanon; Charuta Agashe; Adeeb Rahman; M. Cecilia Berin

doi:10.1007/978-1-0716-2364-0_19

Mass Cytometry Analysis of Whole Blood Response to an Allergen

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

^*Corresponding author for this work

Medicine, Allergy Division

Research output: Chapter in Book/Report/Conference proceeding › Chapter

1 Scopus citations

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 language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	269-280
Number of pages	12
DOIs	https://doi.org/10.1007/978-1-0716-2364-0_19
State	Published - 2022

Publication series

Name	Methods in Molecular Biology
Volume	2506
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Funding

Supported by: U24AI118644 (NIAID).

Keywords

BAT
Basophil
Peanut allergy

ASJC Scopus subject areas

Genetics
Molecular Biology

Access to Document

10.1007/978-1-0716-2364-0_19

Cite this

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title = "Mass Cytometry Analysis of Whole Blood Response to an Allergen",

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.",

keywords = "BAT, Basophil, Peanut allergy",

author = "Daniel Geanon and Charuta Agashe and Adeeb Rahman and Berin, {M. Cecilia}",

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AU - Geanon, Daniel

AU - Agashe, Charuta

AU - Rahman, Adeeb

AU - Berin, M. Cecilia

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Y1 - 2022

N2 - 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.

AB - 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.

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Mass Cytometry Analysis of Whole Blood Response to an Allergen

Abstract

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Keywords

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