Standardizing flow cytometry immunophenotyping analysis from the Human ImmunoPhenotyping Consortium

Finak, Greg, Langweiler, Marc, Jaimes, Maria, Malek, Mehrnoush, Taghiyar, Jafar, Korin, Yael, Raddassi, Khadir, Devine, Lesley, Obermoser, Gerlinde, Pekalski, Marcin L., Pontikos, Nikolas, Diaz, Alain, Heck, Susanne, Villanova, Federica, Terrazzini, Nadia, Kern, Florian, Qian, Yu, Stanton, Rick, Wang, Kui, Brandes, Aaron, Ramey, John, Aghaeepour, Nima, Mosmann, Tim, Scheuermann, Richard H., Reed, Elaine, Palucka, Karolina, Pascual, Virginia, Blomberg, Bonnie B., Nestle, Frank, Nussenblatt, Robert B., Brinkman, Ryan Remy, Gottardo, Raphael, Maecker, Holden and McCoy, J Philip (2016) Standardizing flow cytometry immunophenotyping analysis from the Human ImmunoPhenotyping Consortium. Scientific Reports, 6 20686.1-20686.11. doi:10.1038/srep20686

Author Finak, Greg
Langweiler, Marc
Jaimes, Maria
Malek, Mehrnoush
Taghiyar, Jafar
Korin, Yael
Raddassi, Khadir
Devine, Lesley
Obermoser, Gerlinde
Pekalski, Marcin L.
Pontikos, Nikolas
Diaz, Alain
Heck, Susanne
Villanova, Federica
Terrazzini, Nadia
Kern, Florian
Qian, Yu
Stanton, Rick
Wang, Kui
Brandes, Aaron
Ramey, John
Aghaeepour, Nima
Mosmann, Tim
Scheuermann, Richard H.
Reed, Elaine
Palucka, Karolina
Pascual, Virginia
Blomberg, Bonnie B.
Nestle, Frank
Nussenblatt, Robert B.
Brinkman, Ryan Remy
Gottardo, Raphael
Maecker, Holden
McCoy, J Philip
Title Standardizing flow cytometry immunophenotyping analysis from the Human ImmunoPhenotyping Consortium
Journal name Scientific Reports   Check publisher's open access policy
ISSN 2045-2322
Publication date 2016-02-10
Year available 2016
Sub-type Article (original research)
DOI 10.1038/srep20686
Open Access Status DOI
Volume 6
Start page 20686.1
End page 20686.11
Total pages 12
Place of publication London, United Kingdom
Publisher Nature Publishing Group
Language eng
Subject 1000 General
Abstract Standardization of immunophenotyping requires careful attention to reagents, sample handling, instrument setup, and data analysis, and is essential for successful cross-study and cross-center comparison of data. Experts developed five standardized, eight-color panels for identification of major immune cell subsets in peripheral blood. These were produced as pre-configured, lyophilized, reagents in 96-well plates. We present the results of a coordinated analysis of samples across nine laboratories using these panels with standardized operating procedures (SOPs). Manual gating was performed by each site and by a central site. Automated gating algorithms were developed and tested by the FlowCAP consortium. Centralized manual gating can reduce cross-center variability, and we sought to determine whether automated methods could streamline and standardize the analysis. Within-site variability was low in all experiments, but cross-site variability was lower when central analysis was performed in comparison with site-specific analysis. It was also lower for clearly defined cell subsets than those based on dim markers and for rare populations. Automated gating was able to match the performance of central manual analysis for all tested panels, exhibiting little to no bias and comparable variability. Standardized staining, data collection, and automated gating can increase power, reduce variability, and streamline analysis for immunophenotyping.
Keyword Immunophenotyping
Immune cell subsets
Peripheral blood
Standardized flow cytometry
Data collection
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID 5U19AI090019
R01 EB008400
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: School of Mathematics and Physics
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