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. 2022 Apr 8;3(2):101280.
doi: 10.1016/j.xpro.2022.101280. eCollection 2022 Jun 17.

An optimized protocol for phenotyping human granulocytes by mass cytometry

Nora Vivanco Gonzalez  1 , John-Paul Oliveria  1   2 , Dmitry Tebaykin  1 , Geoffrey T Ivison  1 , Kaori Mukai  1   3 , Mindy M Tsai  1   3 , Luciene Borges  1 , Kari C Nadeau  3   4 , Stephen J Galli  1   3   5 , Albert G Tsai  1 , Sean C Bendall  1
Affiliations

An optimized protocol for phenotyping human granulocytes by mass cytometry

Nora Vivanco Gonzalez et al. STAR Protoc. .

Abstract

Granulocytes encompass diverse roles, from fighting off pathogens to regulating inflammatory processes in allergies. These roles are represented by distinct cellular phenotypes that we captured with mass cytometry (CyTOF). Our protocol enables simultaneous evaluation of human basophils, eosinophils, and neutrophils under homeostasis and upon immune activation by anti-Immunoglobulin E (anti-IgE) or interleukin-3 (IL-3). Granulocyte integrity and detection of protein markers were optimized so that rare granulocyte populations could be deeply characterized by single cell mass cytometry. For complete details on the use and execution of this protocol, please refer to Vivanco Gonzalez et al. (2020).

Keywords: Antibody; Cell isolation; Flow Cytometry/Mass Cytometry; Health Sciences; Immunology; Single Cell.

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Conflict of interest statement

The authors declare no competing interests.

Figures

None
Graphical abstract
Figure 1
Figure 1
Optimization of eosinophil and neutrophil detection (A) Cells were treated with RBC lysis buffer, followed by staining with surface antibodies (including CD16 and CD66b) prior to fixation with PFA as outlined in protocol. (B) Cells were treated with RBC lysis buffer, fixed with PFA, and then stained with surface antibodies. (C) Cells were treated with a commercially available RBC lysis/fix solution prior to staining with surface antibodies.
See this image and copyright information in PMC

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