Standardizing Flow Cytometry Immunophenotyping Analysis from the Human ImmunoPhenotyping Consortium

Greg Finak; Marc Langweiler; Maria Jaimes; Mehrnoush Malek; Jafar Taghiyar; Yael Korin; Khadir Raddassi; Lesley Devine; Gerlinde Obermoser; Marcin L Pekalski; Nikolas Pontikos; Alain Diaz; Susanne Heck; Federica Villanova; Nadia Terrazzini; Florian Kern; Yu Qian; Rick Stanton; Kui Wang; Aaron Brandes; John Ramey; Nima Aghaeepour; Tim Mosmann; Richard H Scheuermann; Elaine Reed; Karolina Palucka; Virginia Pascual; Bonnie B Blomberg; Frank Nestle; Robert B Nussenblatt; Ryan Remy Brinkman; Raphael Gottardo; Holden Maecker; J Philip McCoy

doi:10.1038/srep20686

Back

Standardizing Flow Cytometry Immunophenotyping Analysis from the Human ImmunoPhenotyping Consortium

Journal article

Open access

Peer reviewed

Standardizing Flow Cytometry Immunophenotyping Analysis from the Human ImmunoPhenotyping Consortium

Greg Finak, Marc Langweiler, Maria Jaimes, Mehrnoush Malek, Jafar Taghiyar, Yael Korin, Khadir Raddassi, Lesley Devine, Gerlinde Obermoser, Marcin L Pekalski, …

Scientific reports, Vol.6(1), pp.20686-20686

2016-02-10

DOI: https://doi.org/10.1038/srep20686

PMCID: PMC4748244

PMID: 26861911

Abstract

Laboratories - standards

Leukocytes, Mononuclear - metabolism

Automation

B-Lymphocytes - cytology

Flow Cytometry - methods

Humans

Algorithms

B-Lymphocytes - immunology

T-Lymphocytes - cytology

T-Lymphocytes - metabolism

Flow Cytometry - standards

Leukocytes, Mononuclear - immunology

Immunophenotyping - standards

Immunophenotyping - methods

Leukocytes, Mononuclear - cytology

T-Lymphocytes - immunology

B-Lymphocytes - metabolism

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.

Files and links (1)

url

https://doi.org/10.1038/srep20686View

Published (Version of record) Open

Metrics

26 Record Views

202 Times Cited - Web of Science

See more details

InCites Highlights

These are selected metrics from InCites Benchmarking & Analytics tool, related to this output

Collaboration types: Domestic collaboration; International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.54 Molecular & Cell Biology - Genetics; 1.54.1543 RNA-Seq
Web Of Science research areas: Biochemical Research Methods
ESI research areas: Biology & Biochemistry

UN Sustainable Development Goals (SDGs)

This output has contributed to the advancement of the following goals:

Source: InCites

Details

Title: Standardizing Flow Cytometry Immunophenotyping Analysis from the Human ImmunoPhenotyping Consortium
Creators: Greg Finak - Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, 98109, WA
Marc Langweiler - Hematology Branch, National Institutes of Health, Bethesda, Maryland, USA
Maria Jaimes - BD Biosciences, San Jose, CA, USA
Mehrnoush Malek - Terry Fox Laboratory , British Columbia Cancer Agency, V3J 4W6, Canada
Jafar Taghiyar - Terry Fox Laboratory , British Columbia Cancer Agency, V3J 4W6, Canada
Yael Korin - UCLA Pathology and Laboratory Medicine, Los Angeles, CA
Khadir Raddassi - Dept of Neurology, Yale School of Medicine, New Haven, CT
Lesley Devine - Dept of Neurology, Yale School of Medicine, New Haven, CT
Gerlinde Obermoser - Baylor Institute for Immunology Research, Dallas, TX
Marcin L Pekalski - University of Cambridge, JDRF/Wellcome Trust Diabetes and Inflammation Laboratory, Cambridge Institute for Medical Research, Cambridge, UK
Nikolas Pontikos - University of Cambridge, JDRF/Wellcome Trust Diabetes and Inflammation Laboratory, Cambridge Institute for Medical Research, Cambridge, UK
Alain Diaz - Dept Microbiology &Immunology, University of Miami Miller School of Medicine, Miami, FL
Susanne Heck - Guys and St Thomas Hospital, Guy's Hospital, London, UK
Federica Villanova - Guys and St Thomas Hospital, Guy's Hospital, London, UK
Nadia Terrazzini - School of Pharmacy and Biomolecular Sciences, University of Brighton, Brighton, BN2 4GJ, United Kingdom
Florian Kern - Brighton and Sussex Medical School, Division of Medicine, Brighton, BN1 9PS, United Kingdom
Yu Qian - Department of Informatics, J. Craig Venter Institute, La Jolla, 92037, CA
Rick Stanton - Department of Informatics, J. Craig Venter Institute, La Jolla, 92037, CA
Kui Wang - School of Mathematics and Physics, University of Queensland, Brisbane, Australia
Aaron Brandes - The Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
John Ramey - Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, 98109, WA
Nima Aghaeepour - Baxter Laboratory in Stem Cell Biology, Stanford University, Stanford, California, 94305, USA
Tim Mosmann - University of Rochester Medical Center, School of Medicine and Dentistry, Rochester, 14642, NY
Richard H Scheuermann - Department of Informatics, J. Craig Venter Institute, La Jolla, 92037, CA
Elaine Reed - UCLA Pathology and Laboratory Medicine, Los Angeles, CA
Karolina Palucka - Baylor Institute for Immunology Research, Dallas, TX
Virginia Pascual - Baylor Institute for Immunology Research, Dallas, TX
Bonnie B Blomberg - Dept Microbiology &Immunology, University of Miami Miller School of Medicine, Miami, FL
Frank Nestle - Guys and St Thomas Hospital, Guy's Hospital, London, UK
Robert B Nussenblatt - Laboratory of Immunology, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA
Ryan Remy Brinkman - Department of Medical Genetics, University of British Columbia, Canada
Raphael Gottardo - Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, 98109, WA
Holden Maecker - Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Stanford, 94305, CA
J Philip McCoy - NHLBI Flow Cytometry Core, NIH, Bethesda, 20892, MD
Publication Details: Scientific reports, Vol.6(1), pp.20686-20686
Publisher: England
Grant note: 5U19AI090019 / NIAID NIH HHS R01 EB008400 / NIBIB NIH HHS U19 AI090019 / NIAID NIH HHS Intramural NIH HHS U19 AI089986 / NIAID NIH HHS 100140 / Wellcome Trust
Academic Unit: Miller School of Medicine; Microbiology and Immunology Research
Language: English
Resource Type: Journal article
PMID: 26861911
PMCID: PMC4748244
Record Identifier: 991031577666102976