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High-throughput mass cytometry staining for deep phenotyping of human natural killer cells.
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.
Afiliação
  • Ben Amara A; Immunomonitoring Department, Institut Paoli-Calmettes, 13009 Marseille, France; Team Immunity and Cancer, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM105, 13009 Marseille, France.
  • Rouviere MS; Immunomonitoring Department, Institut Paoli-Calmettes, 13009 Marseille, France; Team Immunity and Cancer, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM105, 13009 Marseille, France.
  • Fattori S; Immunomonitoring Department, Institut Paoli-Calmettes, 13009 Marseille, France; Team Immunity and Cancer, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM105, 13009 Marseille, France.
  • Wlosik J; Immunomonitoring Department, Institut Paoli-Calmettes, 13009 Marseille, France; Team Immunity and Cancer, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM105, 13009 Marseille, France.
  • Gregori E; Centre d'Immunophénomique-Luminy (Ciphe), Inserm US012, CNRS UMS3367, Aix-Marseille University, 13009 Marseille, France.
  • Boucherit N; Immunomonitoring Department, Institut Paoli-Calmettes, 13009 Marseille, France; Team Immunity and Cancer, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM105, 13009 Marseille, France.
  • Bernard PL; Team Immunity and Cancer, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM105, 13009 Marseille, France.
  • Nunès JA; Team Immunity and Cancer, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM105, 13009 Marseille, France.
  • Vey N; Hematology Department, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM 105, 13009 Marseille, France.
  • Luche H; Centre d'Immunophénomique-Luminy (Ciphe), Inserm US012, CNRS UMS3367, Aix-Marseille University, 13009 Marseille, France.
  • Gorvel L; Immunomonitoring Department, Institut Paoli-Calmettes, 13009 Marseille, France; Team Immunity and Cancer, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM105, 13009 Marseille, France. Electronic address: lau
  • Olive D; Immunomonitoring Department, Institut Paoli-Calmettes, 13009 Marseille, France; Team Immunity and Cancer, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM105, 13009 Marseille, France. Electronic address: dan
  • Chretien AS; Immunomonitoring Department, Institut Paoli-Calmettes, 13009 Marseille, France; Team Immunity and Cancer, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068, CNRS, UMR7258, Institut Paoli-Calmettes, Aix-Marseille University, UM105, 13009 Marseille, France. Electronic address: ann
STAR Protoc ; 3(4): 101768, 2022 12 16.
Article em En | MEDLINE | ID: mdl-36269638
ABSTRACT
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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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Leucócitos Mononucleares / Células Matadoras Naturais Tipo de estudo: Guideline Limite: Humans Idioma: En Revista: STAR Protoc Ano de publicação: 2022 Tipo de documento: Article País de afiliação: França
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Leucócitos Mononucleares / Células Matadoras Naturais Tipo de estudo: Guideline Limite: Humans Idioma: En Revista: STAR Protoc Ano de publicação: 2022 Tipo de documento: Article País de afiliação: França