Abstract
These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.
Original language | English |
---|---|
Pages (from-to) | 1457-1973 |
Number of pages | 517 |
Journal | European Journal of Immunology |
Volume | 49 |
Issue number | 10 |
DOIs | |
Publication status | Published - 1 Oct 2019 |
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In: European Journal of Immunology, Vol. 49, No. 10, 01.10.2019, p. 1457-1973.
Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)
AU - Cossarizza, Andrea
AU - Chang, Hyun Dong
AU - Radbruch, Andreas
AU - Acs, Andreas
AU - Adam, Dieter
AU - Adam-Klages, Sabine
AU - Agace, William W.
AU - Aghaeepour, Nima
AU - Akdis, Mübeccel
AU - Allez, Matthieu
AU - Almeida, Larissa Nogueira
AU - Alvisi, Giorgia
AU - Anderson, Graham
AU - Andrä, Immanuel
AU - Annunziato, Francesco
AU - Anselmo, Achille
AU - Bacher, Petra
AU - Baldari, Cosima T.
AU - Bari, Sudipto
AU - Barnaba, Vincenzo
AU - Barros-Martins, Joana
AU - Battistini, Luca
AU - Bauer, Wolfgang
AU - Baumgart, Sabine
AU - Baumgarth, Nicole
AU - Baumjohann, Dirk
AU - Baying, Bianka
AU - Bebawy, Mary
AU - Becher, Burkhard
AU - Beisker, Wolfgang
AU - Benes, Vladimir
AU - Beyaert, Rudi
AU - Blanco, Alfonso
AU - Boardman, Dominic A.
AU - Bogdan, Christian
AU - Borger, Jessica G.
AU - Borsellino, Giovanna
AU - Boulais, Philip E.
AU - Bradford, Jolene A.
AU - Brenner, Dirk
AU - Brinkman, Ryan R.
AU - Brooks, Anna E.S.
AU - Busch, Dirk H.
AU - Büscher, Martin
AU - Bushnell, Timothy P.
AU - Calzetti, Federica
AU - Cameron, Garth
AU - Cammarata, Ilenia
AU - Cao, Xuetao
AU - Cardell, Susanna L.
AU - Casola, Stefano
AU - Cassatella, Marco A.
AU - Cavani, Andrea
AU - Celada, Antonio
AU - Chatenoud, Lucienne
AU - Chattopadhyay, Pratip K.
AU - Chow, Sue
AU - Christakou, Eleni
AU - Čičin-Šain, Luka
AU - Clerici, Mario
AU - Colombo, Federico S.
AU - Cook, Laura
AU - Cooke, Anne
AU - Cooper, Andrea M.
AU - Corbett, Alexandra J.
AU - Cosma, Antonio
AU - Cosmi, Lorenzo
AU - Coulie, Pierre G.
AU - Cumano, Ana
AU - Cvetkovic, Ljiljana
AU - Dang, Van Duc
AU - Dang-Heine, Chantip
AU - Davey, Martin S.
AU - Davies, Derek
AU - De Biasi, Sara
AU - Del Zotto, Genny
AU - Dela Cruz, Gelo Victoriano
AU - Delacher, Michael
AU - Della Bella, Silvia
AU - Dellabona, Paolo
AU - Deniz, Günnur
AU - Dessing, Mark
AU - Di Santo, James P.
AU - Diefenbach, Andreas
AU - Dieli, Francesco
AU - Dolf, Andreas
AU - Dörner, Thomas
AU - Dress, Regine J.
AU - Dudziak, Diana
AU - Dustin, Michael
AU - Dutertre, Charles Antoine
AU - Ebner, Friederike
AU - Eckle, Sidonia B.G.
AU - Edinger, Matthias
AU - Eede, Pascale
AU - Ehrhardt, Götz R.A.
AU - Eich, Marcus
AU - Engel, Pablo
AU - Engelhardt, Britta
AU - Erdei, Anna
AU - Esser, Charlotte
AU - Everts, Bart
AU - Evrard, Maximilien
AU - Falk, Christine S.
AU - Fehniger, Todd A.
AU - Felipo-Benavent, Mar
AU - Ferry, Helen
AU - Feuerer, Markus
AU - Filby, Andrew
AU - Filkor, Kata
AU - Fillatreau, Simon
AU - Follo, Marie
AU - Förster, Irmgard
AU - Foster, John
AU - Foulds, Gemma A.
AU - Frehse, Britta
AU - Frenette, Paul S.
AU - Frischbutter, Stefan
AU - Fritzsche, Wolfgang
AU - Galbraith, David W.
AU - Gangaev, Anastasia
AU - Garbi, Natalio
AU - Gaudilliere, Brice
AU - Gazzinelli, Ricardo T.
AU - Geginat, Jens
AU - Gerner, Wilhelm
AU - Gherardin, Nicholas A.
AU - Ghoreschi, Kamran
AU - Gibellini, Lara
AU - Ginhoux, Florent
AU - Goda, Keisuke
AU - Godfrey, Dale I.
AU - Goettlinger, Christoph
AU - González-Navajas, Jose M.
AU - Goodyear, Carl S.
AU - Gori, Andrea
AU - Grogan, Jane L.
AU - Grummitt, Daryl
AU - Grützkau, Andreas
AU - Haftmann, Claudia
AU - Hahn, Jonas
AU - Hammad, Hamida
AU - Hämmerling, Günter
AU - Hansmann, Leo
AU - Hansson, Goran
AU - Harpur, Christopher M.
AU - Hartmann, Susanne
AU - Hauser, Andrea
AU - Hauser, Anja E.
AU - Haviland, David L.
AU - Hedley, David
AU - Hernández, Daniela C.
AU - Herrera, Guadalupe
AU - Herrmann, Martin
AU - Hess, Christoph
AU - Höfer, Thomas
AU - Hoffmann, Petra
AU - Hogquist, Kristin
AU - Holland, Tristan
AU - Höllt, Thomas
AU - Holmdahl, Rikard
AU - Hombrink, Pleun
AU - Houston, Jessica P.
AU - Hoyer, Bimba F.
AU - Huang, Bo
AU - Huang, Fang Ping
AU - Huber, Johanna E.
AU - Huehn, Jochen
AU - Hundemer, Michael
AU - Hunter, Christopher A.
AU - Hwang, William Y.K.
AU - Iannone, Anna
AU - Ingelfinger, Florian
AU - Ivison, Sabine M.
AU - Jäck, Hans Martin
AU - Jani, Peter K.
AU - Jávega, Beatriz
AU - Jonjic, Stipan
AU - Kaiser, Toralf
AU - Kalina, Tomas
AU - Kamradt, Thomas
AU - Kaufmann, Stefan H.E.
AU - Keller, Baerbel
AU - Ketelaars, Steven L.C.
AU - Khalilnezhad, Ahad
AU - Khan, Srijit
AU - Kisielow, Jan
AU - Klenerman, Paul
AU - Knopf, Jasmin
AU - Koay, Hui Fern
AU - Kobow, Katja
AU - Kolls, Jay K.
AU - Kong, Wan Ting
AU - Kopf, Manfred
AU - Korn, Thomas
AU - Kriegsmann, Katharina
AU - Kristyanto, Hendy
AU - Kroneis, Thomas
AU - Krueger, Andreas
AU - Kühne, Jenny
AU - Kukat, Christian
AU - Kunkel, Désirée
AU - Kunze-Schumacher, Heike
AU - Kurosaki, Tomohiro
AU - Kurts, Christian
AU - Kvistborg, Pia
AU - Kwok, Immanuel
AU - Landry, Jonathan
AU - Lantz, Olivier
AU - Lanuti, Paola
AU - LaRosa, Francesca
AU - Lehuen, Agnès
AU - LeibundGut-Landmann, Salomé
AU - Leipold, Michael D.
AU - Leung, Leslie Y.T.
AU - Levings, Megan K.
AU - Lino, Andreia C.
AU - Liotta, Francesco
AU - Litwin, Virginia
AU - Liu, Yanling
AU - Ljunggren, Hans Gustaf
AU - Lohoff, Michael
AU - Lombardi, Giovanna
AU - Lopez, Lilly
AU - López-Botet, Miguel
AU - Lovett-Racke, Amy E.
AU - Lubberts, Erik
AU - Luche, Herve
AU - Ludewig, Burkhard
AU - Lugli, Enrico
AU - Lunemann, Sebastian
AU - Maecker, Holden T.
AU - Maggi, Laura
AU - Maguire, Orla
AU - Mair, Florian
AU - Mair, Kerstin H.
AU - Mantovani, Alberto
AU - Manz, Rudolf A.
AU - Marshall, Aaron J.
AU - Martínez-Romero, Alicia
AU - Martrus, Glòria
AU - Marventano, Ivana
AU - Maslinski, Wlodzimierz
AU - Matarese, Giuseppe
AU - Mattioli, Anna Vittoria
AU - Maueröder, Christian
AU - Mazzoni, Alessio
AU - McCluskey, James
AU - McGrath, Mairi
AU - McGuire, Helen M.
AU - McInnes, Iain B.
AU - Mei, Henrik E.
AU - Melchers, Fritz
AU - Melzer, Susanne
AU - Mielenz, Dirk
AU - Miller, Stephen D.
AU - Mills, Kingston H.G.
AU - Minderman, Hans
AU - Mjösberg, Jenny
AU - Moore, Jonni
AU - Moran, Barry
AU - Moretta, Lorenzo
AU - Mosmann, Tim R.
AU - Müller, Susann
AU - Multhoff, Gabriele
AU - Muñoz, Luis Enrique
AU - Münz, Christian
AU - Nakayama, Toshinori
AU - Nasi, Milena
AU - Neumann, Katrin
AU - Ng, Lai Guan
AU - Niedobitek, Antonia
AU - Nourshargh, Sussan
AU - Núñez, Gabriel
AU - O'Connor, José Enrique
AU - Ochel, Aaron
AU - Oja, Anna
AU - Ordonez, Diana
AU - Orfao, Alberto
AU - Orlowski-Oliver, Eva
AU - Ouyang, Wenjun
AU - Oxenius, Annette
AU - Palankar, Raghavendra
AU - Panse, Isabel
AU - Pattanapanyasat, Kovit
AU - Paulsen, Malte
AU - Pavlinic, Dinko
AU - Penter, Livius
AU - Peterson, Pärt
AU - Peth, Christian
AU - Petriz, Jordi
AU - Piancone, Federica
AU - Pickl, Winfried F.
AU - Piconese, Silvia
AU - Pinti, Marcello
AU - Pockley, A. Graham
AU - Podolska, Malgorzata Justyna
AU - Poon, Zhiyong
AU - Pracht, Katharina
AU - Prinz, Immo
AU - Pucillo, Carlo E.M.
AU - Quataert, Sally A.
AU - Quatrini, Linda
AU - Quinn, Kylie M.
AU - Radbruch, Helena
AU - Radstake, Tim R.D.J.
AU - Rahmig, Susann
AU - Rahn, Hans Peter
AU - Rajwa, Bartek
AU - Ravichandran, Gevitha
AU - Raz, Yotam
AU - Rebhahn, Jonathan A.
AU - Recktenwald, Diether
AU - Reimer, Dorothea
AU - Reis e Sousa, Caetano
AU - Remmerswaal, Ester B.M.
AU - Richter, Lisa
AU - Rico, Laura G.
AU - Riddell, Andy
AU - Rieger, Aja M.
AU - Robinson, J. Paul
AU - Romagnani, Chiara
AU - Rubartelli, Anna
AU - Ruland, Jürgen
AU - Saalmüller, Armin
AU - Saeys, Yvan
AU - Saito, Takashi
AU - Sakaguchi, Shimon
AU - Sala-de-Oyanguren, Francisco
AU - Samstag, Yvonne
AU - Sanderson, Sharon
AU - Sandrock, Inga
AU - Santoni, Angela
AU - Sanz, Ramon Bellmàs
AU - Saresella, Marina
AU - Sautes-Fridman, Catherine
AU - Sawitzki, Birgit
AU - Schadt, Linda
AU - Scheffold, Alexander
AU - Scherer, Hans U.
AU - Schiemann, Matthias
AU - Schildberg, Frank A.
AU - Schimisky, Esther
AU - Schlitzer, Andreas
AU - Schlosser, Josephine
AU - Schmid, Stephan
AU - Schmitt, Steffen
AU - Schober, Kilian
AU - Schraivogel, Daniel
AU - Schuh, Wolfgang
AU - Schüler, Thomas
AU - Schulte, Reiner
AU - Schulz, Axel Ronald
AU - Schulz, Sebastian R.
AU - Scottá, Cristiano
AU - Scott-Algara, Daniel
AU - Sester, David P.
AU - Shankey, T. Vincent
AU - Silva-Santos, Bruno
AU - Simon, Anna Katharina
AU - Sitnik, Katarzyna M.
AU - Sozzani, Silvano
AU - Speiser, Daniel E.
AU - Spidlen, Josef
AU - Stahlberg, Anders
AU - Stall, Alan M.
AU - Stanley, Natalie
AU - Stark, Regina
AU - Stehle, Christina
AU - Steinmetz, Tobit
AU - Stockinger, Hannes
AU - Takahama, Yousuke
AU - Takeda, Kiyoshi
AU - Tan, Leonard
AU - Tárnok, Attila
AU - Tiegs, Gisa
AU - Toldi, Gergely
AU - Tornack, Julia
AU - Traggiai, Elisabetta
AU - Trebak, Mohamed
AU - Tree, Timothy I.M.
AU - Trotter, Joe
AU - Trowsdale, John
AU - Tsoumakidou, Maria
AU - Ulrich, Henning
AU - Urbanczyk, Sophia
AU - van de Veen, Willem
AU - van den Broek, Maries
AU - van der Pol, Edwin
AU - Van Gassen, Sofie
AU - Van Isterdael, Gert
AU - van Lier, René A.W.
AU - Veldhoen, Marc
AU - Vento-Asturias, Salvador
AU - Vieira, Paulo
AU - Voehringer, David
AU - Volk, Hans Dieter
AU - von Borstel, Anouk
AU - von Volkmann, Konrad
AU - Waisman, Ari
AU - Walker, Rachael V.
AU - Wallace, Paul K.
AU - Wang, Sa A.
AU - Wang, Xin M.
AU - Ward, Michael D.
AU - Ward-Hartstonge, Kirsten A.
AU - Warnatz, Klaus
AU - Warnes, Gary
AU - Warth, Sarah
AU - Waskow, Claudia
AU - Watson, James V.
AU - Watzl, Carsten
AU - Wegener, Leonie
AU - Weisenburger, Thomas
AU - Wiedemann, Annika
AU - Wienands, Jürgen
AU - Wilharm, Anneke
AU - Wilkinson, Robert John
AU - Willimsky, Gerald
AU - Wing, James B.
AU - Winkelmann, Rieke
AU - Winkler, Thomas H.
AU - Wirz, Oliver F.
AU - Wong, Alicia
AU - Wurst, Peter
AU - Yang, Jennie H.M.
AU - Yang, Juhao
AU - Yazdanbakhsh, Maria
AU - Yu, Liping
AU - Yue, Alice
AU - Zhang, Hanlin
AU - Zhao, Yi
AU - Ziegler, Susanne Maria
AU - Zielinski, Christina
AU - Zimmermann, Jakob
AU - Zychlinsky, Arturo
N1 - Funding Information: Ramon Bellmàs Sanz, Jenny Kühne, and Christine S. Falk thank Jana Keil and Kerstin Daemen for excellent technical support. The work was funded by the Germany Research Foundation CRC738/B3 (CSF). Funding Information: Larissa Nogueira Almeida was supported by DFG research grant MA 2273/14‐1. Rudolf A. Manz was supported by the Excellence Cluster “Inflammation at Interfaces” (EXC 306/2). Funding Information: Susanne Hartmann and Friederike Ebner were supported by the German Research Foundation (GRK 2046). Funding Information: Edwin van der Pol and Paola Lanuti acknowledge Aleksandra Gąsecka M.D. for excellent experimental support and Dr. Rienk Nieuwland for textual suggestions. This work was supported by the Netherlands Organisation for Scientific Research – Domain Applied and Engineering Sciences (NWO‐TTW), research program VENI 15924. Funding Information: Hans Minderman was supported by NIH R50CA211108. Funding Information: Work in the laboratory of Dieter Adam is supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—Projektnummer 125440785 – SFB 877, Project B2. Funding Information: Christian Kukat acknowledges support from the ISAC SRL Emerging Leaders program. Funding Information: This work was funded by the Deutsche Forschungsgemeinschaft through the grant TRR130 (project P11 and C03) to Thomas H. Winkler. Funding Information: The Immunology Database and Analysis Portal (ImmPort) system provides an archive of immunology research data generated by investigators mainly funded through the National Institutes of Health (NIH), National Institute of Allergy and Infectious Diseases (NIAID), and Division of Allergy, Immunology, and Transplantation (DAIT). It is an extensive data warehouse containing an integration of experimental and clinical trial data generated by dozens of assay types, including 131 FCM and 24 CyTOF data sets. In addition, the ImmPort system also provides data analysis tools and it contains implicit knowledge and ‘‘best practices’’ for clinical and genomic studies in the form of ∼50 templates for data deposition, management, and dissemination. ImmPort has been developed under the Bioinformatics Integration Support Contract (BISC) by the Northrop Grumman Information Technology Health Solutions team for the NIH NIAID/DAIT. If your research funding comes from this source and you are generating immunology data, you should deposit it in ImmPort. Immport's support for different data types can be another reason to choose it if you are generating FCM data as well as data from different types of assays. A (free) registration and approval by DAIT is required in order to deposit and access data from ImmPort. A potential drawback of using this resource is that there can be a long delay in publication of deposited datasets. Funding Information: The work by the Mei laboratory was supported by German Research Foundation Grant ME 3644/5‐1 and TRR130 TP24, the German Rheumatism Research Centre Berlin, European Union Innovative Medicines Initiative ‐ Joint Undertaking ‐ RTCure Grant Agreement 777357, the Else Kröner‐Fresenius‐Foundation, German Federal Ministry of Education and Research e:Med sysINFLAME Program Grant 01ZX1306B and KMU‐innovativ “InnoCyt”, and the Leibniz Science Campus for Chronic Inflammation ( http://www.chronische-entzuendung.org ). Funding Information: Petra Hoffmann, Andrea Hauser, and Matthias Edinger thank BD Biosciences®, San José, CA, USA, and SKAN AG, Bale, Switzerland for fruitful cooperation during the development, construction, and installation of the GMP‐compliant cell sorting equipment and the Bavarian Immune Therapy Network (BayImmuNet) for financial support. Funding Information: John Trowsdale received funding from the European Research Council under the European Union's Horizon 2020 research and innovation program (Grant Agreement 695551). Publisher Copyright: © 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
PY - 2019/10/1
Y1 - 2019/10/1
N2 - These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.
AB - These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.
UR - http://www.scopus.com/inward/record.url?scp=85073598520&partnerID=8YFLogxK
UR - https://pubmed.ncbi.nlm.nih.gov/31633216
U2 - 10.1002/eji.201970107
DO - 10.1002/eji.201970107
M3 - Article
C2 - 31633216
AN - SCOPUS:85073598520
SN - 0014-2980
VL - 49
SP - 1457
EP - 1973
JO - European Journal of Immunology
JF - European Journal of Immunology
IS - 10
ER -