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Comparison of CyTOF assays across sites: Results of a six-center pilot study.
Leipold, Michael D; Obermoser, Gerlinde; Fenwick, Craig; Kleinstuber, Katja; Rashidi, Narges; McNevin, John P; Nau, Allison N; Wagar, Lisa E; Rozot, Virginie; Davis, Mark M; DeRosa, Stephen; Pantaleo, Giuseppe; Scriba, Thomas J; Walker, Bruce D; Olsen, Lars R; Maecker, Holden T.
Afiliación
  • Leipold MD; Human Immune Monitoring Center, Stanford University, Stanford, CA, USA. Electronic address: mleipold@stanford.edu.
  • Obermoser G; Human Immune Monitoring Center, Stanford University, Stanford, CA, USA.
  • Fenwick C; Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland.
  • Kleinstuber K; Ragon Institute of MGH, MIT, and Harvard, Cambridge, MA, USA.
  • Rashidi N; Ragon Institute of MGH, MIT, and Harvard, Cambridge, MA, USA.
  • McNevin JP; Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
  • Nau AN; Institute for Transplantation and Immunity, Stanford University, Stanford, CA, USA.
  • Wagar LE; Institute for Transplantation and Immunity, Stanford University, Stanford, CA, USA.
  • Rozot V; South African Tuberculosis Vaccine Initiative, Institute of Infectious Diseases and Molecular Medicine, Division of Immunology, Department of Pathology, University of Cape Town, Cape Town, South Africa.
  • Davis MM; Institute for Transplantation and Immunity, Stanford University, Stanford, CA, USA.
  • DeRosa S; Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
  • Pantaleo G; Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland.
  • Scriba TJ; South African Tuberculosis Vaccine Initiative, Institute of Infectious Diseases and Molecular Medicine, Division of Immunology, Department of Pathology, University of Cape Town, Cape Town, South Africa.
  • Walker BD; Ragon Institute of MGH, MIT, and Harvard, Cambridge, MA, USA.
  • Olsen LR; DTU Bioinformatics, Technical University of Denmark, Copenhagen, Denmark.
  • Maecker HT; Human Immune Monitoring Center, Stanford University, Stanford, CA, USA.
J Immunol Methods ; 453: 37-43, 2018 02.
Article en En | MEDLINE | ID: mdl-29174717
For more than five years, high-dimensional mass cytometry has been employed to study immunology. However, these studies have typically been performed in one laboratory on one or few instruments. We present the results of a six-center study using healthy control human peripheral blood mononuclear cells (PBMCs) and commercially available reagents to test the intra-site and inter-site variation of mass cytometers and operators. We used prestained controls generated by the primary center as a reference to compare against samples stained at each individual center. Data were analyzed at the primary center, including investigating the effects of two normalization methods. All six sites performed similarly, with CVs for both Frequency of Parent and median signal intensity (MSI) values<30%. Increased background was seen when using the premixed antibody cocktail aliquots at each site, suggesting that cocktails are best made fresh. Both normalization methods tested performed adequately for normalizing MSI values between centers. Clustering algorithms revealed slight differences between the prestained and the sites-stained samples, due mostly to the increased background of a few antibodies. Therefore, we believe that multicenter mass cytometry assays are feasible.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espectrometría de Masas / Leucocitos Mononucleares / Citometría de Flujo Tipo de estudio: Clinical_trials Límite: Humans Idioma: En Revista: J Immunol Methods Año: 2018 Tipo del documento: Article Pais de publicación: Países Bajos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espectrometría de Masas / Leucocitos Mononucleares / Citometría de Flujo Tipo de estudio: Clinical_trials Límite: Humans Idioma: En Revista: J Immunol Methods Año: 2018 Tipo del documento: Article Pais de publicación: Países Bajos