Semi-automated and standardized cytometric procedures for multi-panel and multi-parametric whole blood immunophenotyping
Abstract Immunophenotyping by multi-parametric flow cytometry is the cornerstone technology for enumeration and characterization of immune cell populations in health and disease. Standardized procedures are essential to allow for inter-individual comparisons in the context of population based or cli...
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| Veröffentlicht in: | Clinical immunology (Orlando, Fla.) Fla.), 2015-04, Vol.157 (2), p.261-276 |
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| creator | Hasan, Milena Beitz, Benoit Rouilly, Vincent Libri, Valentina Urrutia, Alejandra Duffy, Darragh Cassard, Lydie Di Santo, James P Mottez, Estelle Quintana-Murci, Lluis Albert, Matthew L Rogge, Lars |
| description | Abstract Immunophenotyping by multi-parametric flow cytometry is the cornerstone technology for enumeration and characterization of immune cell populations in health and disease. Standardized procedures are essential to allow for inter-individual comparisons in the context of population based or clinical studies. Herein we report the approach taken by the Milieu Intérieur Consortium, highlighting the standardized and automated procedures used for immunophenotyping of human whole blood samples. We optimized eight-color antibody panels and procedures for staining and lysis of whole blood samples, and implemented pre-analytic steps with a semi-automated workflow using a robotic system. We report on four panels that were designed to enumerate and phenotype major immune cell populations (PMN, T, B, NK cells, monocytes and DC). This work establishes a foundation for defining reference values in healthy donors. Our approach provides robust protocols for affordable, semi-automated eight-color cytometric immunophenotyping that can be used in population-based studies and clinical trial settings. |
| doi_str_mv | 10.1016/j.clim.2014.12.008 |
| format | Article |
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Standardized procedures are essential to allow for inter-individual comparisons in the context of population based or clinical studies. Herein we report the approach taken by the Milieu Intérieur Consortium, highlighting the standardized and automated procedures used for immunophenotyping of human whole blood samples. We optimized eight-color antibody panels and procedures for staining and lysis of whole blood samples, and implemented pre-analytic steps with a semi-automated workflow using a robotic system. We report on four panels that were designed to enumerate and phenotype major immune cell populations (PMN, T, B, NK cells, monocytes and DC). This work establishes a foundation for defining reference values in healthy donors. Our approach provides robust protocols for affordable, semi-automated eight-color cytometric immunophenotyping that can be used in population-based studies and clinical trial settings.</description><identifier>ISSN: 1521-6616</identifier><identifier>EISSN: 1521-7035</identifier><identifier>DOI: 10.1016/j.clim.2014.12.008</identifier><identifier>PMID: 25572534</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Allergy and Immunology ; Antigens, CD - immunology ; Automation ; Automation, Laboratory - methods ; B-Lymphocytes ; Cytometry ; Dendritic Cells ; Flow Cytometry - methods ; Humans ; Immunology ; Immunophenotyping ; Immunophenotyping - methods ; Killer Cells, Natural ; Life Sciences ; Monocytes ; Neutrophils ; Specimen Handling - methods ; Standardization ; T-Lymphocytes ; Whole blood</subject><ispartof>Clinical immunology (Orlando, Fla.), 2015-04, Vol.157 (2), p.261-276</ispartof><rights>The Authors</rights><rights>2015 The Authors</rights><rights>Copyright © 2015. Published by Elsevier Inc.</rights><rights>Attribution - NonCommercial - NoDerivatives</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c596t-13ed6a13753411daa594b8d04e5d09755270ad5a44549132b634702e2165e1033</citedby><cites>FETCH-LOGICAL-c596t-13ed6a13753411daa594b8d04e5d09755270ad5a44549132b634702e2165e1033</cites><orcidid>0000-0003-2429-6320 ; 0000-0001-8373-041X ; 0000-0002-8875-2308 ; 0000-0002-1691-4531 ; 0000-0002-0879-7099</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S1521661614002861$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>230,314,776,780,881,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25572534$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://pasteur.hal.science/pasteur-01380961$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Hasan, Milena</creatorcontrib><creatorcontrib>Beitz, Benoit</creatorcontrib><creatorcontrib>Rouilly, Vincent</creatorcontrib><creatorcontrib>Libri, Valentina</creatorcontrib><creatorcontrib>Urrutia, Alejandra</creatorcontrib><creatorcontrib>Duffy, Darragh</creatorcontrib><creatorcontrib>Cassard, Lydie</creatorcontrib><creatorcontrib>Di Santo, James P</creatorcontrib><creatorcontrib>Mottez, Estelle</creatorcontrib><creatorcontrib>Quintana-Murci, Lluis</creatorcontrib><creatorcontrib>Albert, Matthew L</creatorcontrib><creatorcontrib>Rogge, Lars</creatorcontrib><creatorcontrib>Consortium</creatorcontrib><creatorcontrib>for The</creatorcontrib><creatorcontrib>Milieu Intérieur Consortium</creatorcontrib><title>Semi-automated and standardized cytometric procedures for multi-panel and multi-parametric whole blood immunophenotyping</title><title>Clinical immunology (Orlando, Fla.)</title><addtitle>Clin Immunol</addtitle><description>Abstract Immunophenotyping by multi-parametric flow cytometry is the cornerstone technology for enumeration and characterization of immune cell populations in health and disease. Standardized procedures are essential to allow for inter-individual comparisons in the context of population based or clinical studies. Herein we report the approach taken by the Milieu Intérieur Consortium, highlighting the standardized and automated procedures used for immunophenotyping of human whole blood samples. We optimized eight-color antibody panels and procedures for staining and lysis of whole blood samples, and implemented pre-analytic steps with a semi-automated workflow using a robotic system. We report on four panels that were designed to enumerate and phenotype major immune cell populations (PMN, T, B, NK cells, monocytes and DC). This work establishes a foundation for defining reference values in healthy donors. 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Beitz, Benoit ; Rouilly, Vincent ; Libri, Valentina ; Urrutia, Alejandra ; Duffy, Darragh ; Cassard, Lydie ; Di Santo, James P ; Mottez, Estelle ; Quintana-Murci, Lluis ; Albert, Matthew L ; Rogge, Lars</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c596t-13ed6a13753411daa594b8d04e5d09755270ad5a44549132b634702e2165e1033</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Allergy and Immunology</topic><topic>Antigens, CD - immunology</topic><topic>Automation</topic><topic>Automation, Laboratory - methods</topic><topic>B-Lymphocytes</topic><topic>Cytometry</topic><topic>Dendritic Cells</topic><topic>Flow Cytometry - methods</topic><topic>Humans</topic><topic>Immunology</topic><topic>Immunophenotyping</topic><topic>Immunophenotyping - methods</topic><topic>Killer Cells, Natural</topic><topic>Life Sciences</topic><topic>Monocytes</topic><topic>Neutrophils</topic><topic>Specimen Handling - methods</topic><topic>Standardization</topic><topic>T-Lymphocytes</topic><topic>Whole blood</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hasan, Milena</creatorcontrib><creatorcontrib>Beitz, Benoit</creatorcontrib><creatorcontrib>Rouilly, Vincent</creatorcontrib><creatorcontrib>Libri, Valentina</creatorcontrib><creatorcontrib>Urrutia, Alejandra</creatorcontrib><creatorcontrib>Duffy, Darragh</creatorcontrib><creatorcontrib>Cassard, Lydie</creatorcontrib><creatorcontrib>Di Santo, James P</creatorcontrib><creatorcontrib>Mottez, Estelle</creatorcontrib><creatorcontrib>Quintana-Murci, Lluis</creatorcontrib><creatorcontrib>Albert, Matthew L</creatorcontrib><creatorcontrib>Rogge, Lars</creatorcontrib><creatorcontrib>Consortium</creatorcontrib><creatorcontrib>for The</creatorcontrib><creatorcontrib>Milieu Intérieur Consortium</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><jtitle>Clinical immunology (Orlando, Fla.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hasan, Milena</au><au>Beitz, Benoit</au><au>Rouilly, Vincent</au><au>Libri, Valentina</au><au>Urrutia, Alejandra</au><au>Duffy, Darragh</au><au>Cassard, Lydie</au><au>Di Santo, James P</au><au>Mottez, Estelle</au><au>Quintana-Murci, Lluis</au><au>Albert, Matthew L</au><au>Rogge, Lars</au><aucorp>Consortium</aucorp><aucorp>for The</aucorp><aucorp>Milieu Intérieur Consortium</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Semi-automated and standardized cytometric procedures for multi-panel and multi-parametric whole blood immunophenotyping</atitle><jtitle>Clinical immunology (Orlando, Fla.)</jtitle><addtitle>Clin Immunol</addtitle><date>2015-04-01</date><risdate>2015</risdate><volume>157</volume><issue>2</issue><spage>261</spage><epage>276</epage><pages>261-276</pages><issn>1521-6616</issn><eissn>1521-7035</eissn><abstract>Abstract Immunophenotyping by multi-parametric flow cytometry is the cornerstone technology for enumeration and characterization of immune cell populations in health and disease. 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| ispartof | Clinical immunology (Orlando, Fla.), 2015-04, Vol.157 (2), p.261-276 |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Allergy and Immunology Antigens, CD - immunology Automation Automation, Laboratory - methods B-Lymphocytes Cytometry Dendritic Cells Flow Cytometry - methods Humans Immunology Immunophenotyping Immunophenotyping - methods Killer Cells, Natural Life Sciences Monocytes Neutrophils Specimen Handling - methods Standardization T-Lymphocytes Whole blood |
| title | Semi-automated and standardized cytometric procedures for multi-panel and multi-parametric whole blood immunophenotyping |
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