High-throughput mass cytometry staining for deep phenotyping of human natural killer cells

This protocol details the step-by-step procedure for in-depth immune phenotyping of peripheral blood natural killer (NK) cells from clinical samples by mass cytometry. The protocol consists of three main steps: PBMC incubation with a mix of metal-conjugated antibodies for extracellular phenotyping f...

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Detalles Bibliográficos
Autores principales: Ben Amara, Amira, Rouviere, Marie-Sarah, Fattori, Stéphane, Wlosik, Julia, Gregori, Emilie, Boucherit, Nicolas, Bernard, Pierre-Louis, Nunès, Jacques A., Vey, Norbert, Luche, Herve, Gorvel, Laurent, Olive, Daniel, Chretien, Anne-Sophie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9589031/
https://www.ncbi.nlm.nih.gov/pubmed/36269638
http://dx.doi.org/10.1016/j.xpro.2022.101768
Descripción
Sumario:This protocol details the step-by-step procedure for in-depth immune phenotyping of peripheral blood natural killer (NK) cells from clinical samples by mass cytometry. The protocol consists of three main steps: PBMC incubation with a mix of metal-conjugated antibodies for extracellular phenotyping followed by fixation, permeabilization and incubation with a mix of metal-conjugated antibodies for staining of intracellular proteins, and sample acquisition on a mass cytometer. High-dimensional analysis enables the visualization of NK cell subsets and their phenotypical characteristics. For complete details on the use and execution of this protocol, please refer to Chretien et al. (2021).

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