Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)
Andrea Cossarizza(University of Modena and Reggio Emilia), Hyun‐Dong Chang(Leibniz Association), Andreas Radbruch(Leibniz Association), Andreas Acs(Friedrich-Alexander-Universität Erlangen-Nürnberg), Dieter Adam(Christian-Albrechts-Universität zu Kiel), Sabine Adam‐Klages, William W. Agace(Lund University), Nima Aghaeepour(Stanford Medicine), Mübeccel Akdiş(University of Zurich), Matthieu Allez(Inserm), Larissa N. Almeida(Institute for Integrative and Experimental Genomics), Giorgia Alvisi(Humanitas University), Graham Anderson(University of Birmingham), Immanuel Andrä(Technical University of Munich), Francesco Annunziato(University of Florence), Achille Anselmo(Humanitas University), Petra Bächer(Christian-Albrechts-Universität zu Kiel), Cosima T. Baldari(University of Siena), Sudipto Bari(Duke-NUS Medical School), Vincenzo Barnaba(Italian Institute of Technology), Joana Barros‐Martins(Medizinische Hochschule Hannover), Luca Battistini(Fondazione Santa Lucia), Wolfgang Bauer(Medical University of Vienna), Sabine Baumgart(Leibniz Association), Nicole Baumgarth(University of California, Davis), Dirk Baumjohann(Urologische Klinik München), Bianka Baying(European Molecular Biology Laboratory), Mary Bebawy(University of Technology Sydney), Burkhard Becher(University of Zurich), Wolfgang Beisker(Helmholtz Zentrum München), Vladimı́r Beneš(European Molecular Biology Laboratory), Rudi Beyaert(Ghent University), Alfonso Blanco(University College Dublin), Dominic A. Boardman(University of British Columbia), Christian Bogdan(Friedrich-Alexander-Universität Erlangen-Nürnberg), Jessica G Borger(Monash University), Giovanna Borsellino(Fondazione Santa Lucia), Philip E. Boulais(Albert Einstein College of Medicine), Jolene A. Bradford(Thermo Fisher Scientific (United States)), Dirk Brenner(University of Southern Denmark), Ryan R. Brinkman(University of British Columbia), Anna E. S. Brooks(University of Auckland), Dirk H. Busch(German Center for Infection Research), Martin Büscher(Miltenyi Biotec (Germany)), Timothy Bushnell(University of Rochester Medical Center), Federica Calzetti(University of Verona), Garth Cameron(The University of Melbourne), Ilenia Cammarata(Sapienza University of Rome), Xuetao Cao(Nankai University), Susanna Cardell(University of Gothenburg), Stefano Casola(IFOM), Marco A. Cassatella(University of Verona), Andrea Cavani(National Institute of Health), Antonio Celada(Universitat de Barcelona), Lucienne Chatenoud(Délégation Paris 5), Pratip K. Chattopadhyay(NYU Langone Health), Sue Chow(Princess Margaret Cancer Centre), Eleni Christakou(National Health Service), Luka Čičin‐Šain(Helmholtz Centre for Infection Research), Mario Clerici(University of Milan), Federico Colombo(Humanitas University), Laura Cook(University of British Columbia), Anne Cooke(University of Cambridge), Andrea M. Cooper(University of Leicester), Alexandra J. Corbett(The University of Melbourne), Antonio Cosma(Luxembourg Institute of Health), Lorenzo Cosmi(University of Florence), Pierre G. Coulie(de Duve Institute), Ana Cumano(Institut Pasteur), Ljiljana Cvetković(Friedrich-Alexander-Universität Erlangen-Nürnberg), Van Duc Dang(Leibniz Association), Chantip Dang‐Heine(Berlin Institute of Health at Charité - Universitätsmedizin Berlin), Martin S. Davey(Australian Research Council), Derek Davies(The Francis Crick Institute), Sara De Biasi(University of Modena and Reggio Emilia), Genny Del Zotto(Istituto Giannina Gaslini), Gelo Victoriano Dela Cruz(University of Copenhagen), Michael Delacher(University Hospital Regensburg), Silvia Della Bella(University of Milan), Paolo Dellabona(Vita-Salute San Raffaele University), Günnur Deniz(HUN-REN Institute of Experimental Medicine), Mark C. Dessing(Sony (United Kingdom)), James P. Di Santo(Inserm), Andreas Diefenbach(Leibniz Association), Francesco Dieli(University of Palermo), Andreas Dolf(University of Bonn), Thomas Dörner(Leibniz Association), Regine J. Dress(Agency for Science, Technology and Research), Diana Dudziak(Friedrich-Alexander-Universität Erlangen-Nürnberg), Michael L. Dustin(University of Oxford), Charles‐Antoine Dutertre(Agency for Science, Technology and Research), Friederike Ebner(Freie Universität Berlin), Sidonia B. G. Eckle(The University of Melbourne), Matthias Edinger(University Hospital Regensburg), Pascale Eede(Humboldt-Universität zu Berlin), Götz R. A. Ehrhardt(University of Toronto), Marcus Eich(Heidelberg Institute for Stem Cell Technology and Experimental Medicine), Pablo Engel(Barcelona Biomedical Research Park), Britta Engelhardt(University of Bern), Anna Erdei(Eötvös Loránd University), Charlotte Esser(Leibniz Association), Bart Everts(Leiden University Medical Center), Maximilien Evrard(Agency for Science, Technology and Research), Christine S. Falk(Medizinische Hochschule Hannover), Todd A. Fehniger(Washington University in St. Louis), Mar Felipo‐Benavent(Centro de Investigacion Principe Felipe), Helen Ferry(University of Oxford), Markus Feuerer(University Hospital Regensburg), Andrew Filby(Newcastle University), Kata Filkor(University of Szeged), Simon Fillatreau(Hôpital Necker-Enfants Malades), Marie Follo(University of Freiburg), Irmgard Förster(University of Bonn), John R. Foster(Owl Biomedical (United States)), Gemma A. Foulds(Nottingham Trent University), Britta Frehse(Institute for Integrative and Experimental Genomics), Paul S. Frenette(Albert Einstein College of Medicine), Stefan Frischbutter(Leibniz Association), Wolfgang Fritzsche(Leibniz Institute of Photonic Technology), David W. Galbraith(University of Arizona), Anastasia Gangaev(The Netherlands Cancer Institute), Natalio Garbi(University of Bonn), Brice Gaudillière(Stanford Medicine), Ricardo T. Gazzinelli(University of Massachusetts Chan Medical School), Jens Geginat(Istituto Nazionale Genetica Molecolare), Wilhelm Gerner(University of Veterinary Medicine Vienna), Nicholas A. Gherardin(The University of Melbourne), Kamran Ghoreschi(Charité - Universitätsmedizin Berlin), Lara Gibellini(University of Modena and Reggio Emilia), Florent Ginhoux(Agency for Science, Technology and Research), Keisuke Goda(University of California, Los Angeles), Dale I. Godfrey(The University of Melbourne), Christoph Goettlinger(University of Cologne), José M. González‐Navajas(Centre for Biomedical Network Research on Rare Diseases), Carl S. Goodyear(Wellcome Centre for Molecular Parasitology), Andrea Gori(University of Milan), Jane L. Grogan(Cancer Research Institute), Daryl Grummitt(Owl Biomedical (United States)), Andreas Grützkau(Leibniz Association), Claudia Haftmann(University of Zurich), Jonas Hahn(Friedrich-Alexander-Universität Erlangen-Nürnberg), Hamida Hammad(Ablynx (Belgium)), Günter J. Hämmerling(German Cancer Research Center), Leo Hansmann(German Cancer Society), Göran K. Hansson(Karolinska University Hospital), Christopher M. Harpur(Murdoch Children's Research Institute), Susanne Hartmann(Freie Universität Berlin), Andrea Hauser(University Hospital Regensburg), Anja E. Hauser(University Hospital Regensburg), David L. Haviland(Houston Methodist), David W. Hedley(Princess Margaret Cancer Centre), Daniela Hernandez(Leibniz Association), Guadalupe Herrera(Universitat de València), Martin Herrmann(Friedrich-Alexander-Universität Erlangen-Nürnberg), Christoph Hess(University of Cambridge), Thomas Höfer(German Cancer Research Center), Petra Hoffmann(University Hospital Regensburg), Kristin A. Hogquist(University of Minnesota), Tristan Holland(University of Bonn), Thomas Höllt(Leiden University Medical Center), Rikard Holmdahl(Karolinska Institutet), Pleun Hombrink(Sanquin), Jessica P. Houston(New Mexico State University), Bimba F. Hoyer(University Hospital Schleswig-Holstein), Bo Huang(Chinese Academy of Medical Sciences & Peking Union Medical College), Fang‐Ping Huang(Shenzhen University), Johanna E. Huber(Urologische Klinik München), Jochen Huehn(Helmholtz Centre for Infection Research), Michael Hundemer(Heidelberg University), Christopher A. Hunter(University of Pennsylvania), William Ying Khee Hwang(Singapore General Hospital), Anna Iannone(University of Modena and Reggio Emilia), Florian Ingelfinger(University of Zurich), Sabine Ivison(University of British Columbia), Hans‐Martin Jäck(Friedrich-Alexander-Universität Erlangen-Nürnberg), Péter K. Jani(Leibniz Association), Beatriz Jávega(Universitat de València), Stipan Jonjić(University of Rijeka), Toralf Kaiser(Leibniz Association), Tomáš Kalina(Charles University), Thomas Kamradt(Jena University Hospital), Stefan H. E. Kaufmann(Max Planck Institute for Infection Biology), Baerbel Keller(University of Freiburg), Steven L. C. Ketelaars(The Netherlands Cancer Institute), Ahad Khalilnezhad(Agency for Science, Technology and Research), Srijit Khan(University of Toronto), Jan Kisielow(ETH Zurich), Paul Klenerman(University of Oxford), Jasmin Knopf(Friedrich-Alexander-Universität Erlangen-Nürnberg), Hui‐Fern Koay(The University of Melbourne), Katja Kobow(Universitätsklinikum Erlangen), Jay K. Kolls(Tulane University), Wan Ting Kong(Agency for Science, Technology and Research), Manfred Köpf(ETH Zurich), Thomas Korn(Technical University of Munich), Katharina Kriegsmann(Heidelberg University), Hendy Kristyanto(Leiden University Medical Center), Thomas Kroneis(Medical University of Graz), Andreas Krueger(Goethe University Frankfurt), Jenny F. Kühne(Medizinische Hochschule Hannover), Christian Kukat(Max Planck Institute for Biology of Ageing), Désirée Kunkel(Berlin-Brandenburger Centrum für Regenerative Therapien), Heike Kunze‐Schumacher(Goethe University Frankfurt), Tomohiro Kurosaki(The University of Osaka), Christian Kurts(University of Bonn), Pia Kvistborg(The Netherlands Cancer Institute), Immanuel Kwok(Agency for Science, Technology and Research), Jonathan J. M. Landry(European Molecular Biology Laboratory), Olivier Lantz(Inserm), Paola Lanuti(University of Chieti-Pescara), Francesca Larosa(Don Carlo Gnocchi Foundation), Agnès Lehuen(Centre National de la Recherche Scientifique), Salomé LeibundGut‐Landmann(University of Zurich), Michael D. Leipold(Institute of Infection and Immunity), Leslie Y. T. Leung(University of Toronto), Megan K. Levings(University of British Columbia), Andreia C. Lino(Leibniz Association), Francesco Liotta(University of Florence), Virginia Litwin(Caprion (Canada)), Yanling Liu(University of Toronto), Hans‐Gustaf Ljunggren(Karolinska Institutet), Michael Lohoff(Philipps University of Marburg), Giovanna Lombardi(King's College London), Lilly Lopez(Beckman Coulter Foundation), Miguel López‐Botet(Universitat Pompeu Fabra), Amy E. Lovett‐Racke(Institute of Infection and Immunity), Erik Lubberts(Erasmus MC), Hervé Luche(Centre National de la Recherche Scientifique), Burkhard Ludewig(Kantonsspital St. Gallen), Enrico Lugli(Humanitas University), Sebastian Lunemann(Leibniz Institute of Virology (LIV)), Holden T. Maecker(Institute of Infection and Immunity), Laura Maggi(University of Florence), Orla Maguire(Roswell Park Comprehensive Cancer Center), Florian Mair(Cape Town HVTN Immunology Laboratory / Hutchinson Centre Research Institute of South Africa), Kerstin H. Mair(University of Veterinary Medicine Vienna), Alberto Mantovani(Humanitas University), Rudolf A. Manz(Institute for Integrative and Experimental Genomics), Aaron J. Marshall(University of Manitoba), Alicia Martínez‐Romero(Leitat Technological Center), Glòria Martrus(Leibniz Institute of Virology (LIV)), Ivana Marventano(Don Carlo Gnocchi Foundation), Włodzimierz Maśliński(National Institute of Geriatrics, Rheumatology and Rehabilitation), Giuseppe Matarese(Institute for Experimental Endocrinology and Oncology), Anna Vittoria Mattioli(University of Modena and Reggio Emilia), Christian Maueröder(Ghent University), Alessio Mazzoni(University of Florence), James McCluskey(The University of Melbourne), Mairi McGrath(Leibniz Association), Helen M. McGuire(The University of Sydney), Iain B. McInnes(Wellcome Centre for Molecular Parasitology), Henrik E. Mei(Leibniz Association), Fritz Melchers(Leibniz Association), Susanne Melzer(Leipzig University), Dirk Mielenz(Friedrich-Alexander-Universität Erlangen-Nürnberg), Stephen D. Miller(Northwestern University), Kingston H. G. Mills(Trinity College Dublin), Hans Minderman(Roswell Park Comprehensive Cancer Center), Jenny Mjösberg(Linköping University), Jonni S. Moore(University of Pennsylvania), Barry Moran(Trinity College Dublin), Lorenzo Moretta(Bambino Gesù Children's Hospital), Tim R. Mosmann(University of Rochester Medical Center), Susann Müller(Helmholtz Centre for Environmental Research), Gabriele Multhoff(TUM Klinikum), Luis E. Muñoz(Friedrich-Alexander-Universität Erlangen-Nürnberg), Christian Münz(University of Zurich), Toshinori Nakayama(Chiba University), Milena Nasi(University of Modena and Reggio Emilia), Katrin Neumann(Universität Hamburg), Lai Guan Ng(Agency for Science, Technology and Research), Antonia Niedobitek(Leibniz Association), Sussan Nourshargh(Queen Mary University of London), Gabriel Núñez(University of Michigan), José‐Enrique O’Connor(Universitat de València), Aaron Ochel(Universität Hamburg), Anna E. Oja(Sanquin), Diana Ordonez(European Molecular Biology Laboratory), Alberto Órfão(Universidad de Salamanca), Eva Orlowski‐Oliver(Burnet Institute), Wenjun Ouyang(Amgen (United States)), Annette Oxenius(ETH Zurich), Raghavendra Palankar(Universitätsmedizin Greifswald), Isabel Panse(Leibniz Association), Kovit Pattanapanyasat(Siriraj Hospital), Malte Paulsen(European Molecular Biology Laboratory), Dinko Pavlinić(European Molecular Biology Laboratory), Livius Penter(Charité - Universitätsmedizin Berlin), Pärt Peterson(University of Tartu), Christian Peth(Miltenyi Biotec (Germany)), Jordi Pétriz(Universitat Autònoma de Barcelona), Federica Piancone(Don Carlo Gnocchi Foundation), Winfried F. Pickl(Medical University of Vienna), Silvia Piconese(Istituto Pasteur), Marcello Pinti(University of Modena and Reggio Emilia), A. Graham Pockley(PhytoCeutica (United States)), Malgorzata J. Podolska(Friedrich-Alexander-Universität Erlangen-Nürnberg), Zhiyong Poon(Singapore General Hospital), Katharina Pracht(Friedrich-Alexander-Universität Erlangen-Nürnberg), Immo Prinz(Medizinische Hochschule Hannover), Carlo Pucillo(University of Udine), Sally A. Quataert(University of Rochester Medical Center), Linda Quatrini(Bambino Gesù Children's Hospital), Kylie M. Quinn(Monash University), Helena Radbruch(Humboldt-Universität zu Berlin), Tim Radstake(Utrecht University), Susann Rahmig(Leibniz Institute on Aging - Fritz Lipmann Institute (FLI)), Hans-Peter Rahn(Max Delbrück Center), Bartek Rajwa(Purdue University West Lafayette), Gevitha Ravichandran(Universität Hamburg), Yotam Raz(Groene Hart Ziekenhuis), Jonathan A. Rebhahn(University of Rochester Medical Center), Diether Recktenwald(Desert Research Institute), Dorothea Reimer(Friedrich-Alexander-Universität Erlangen-Nürnberg), Caetano Reis e Sousa(The Francis Crick Institute), Ester B. M. Remmerswaal(Amsterdam University Medical Centers), Lisa Richter(Ludwig-Maximilians-Universität München), Laura G. Rico(Universitat Autònoma de Barcelona), Andrew Riddell(The Francis Crick Institute), Aja M. Rieger(University of Alberta), J. Paul Robinson(Purdue University West Lafayette), Chiara Romagnani(Leibniz Association), Anna Rubartelli(Ospedale Policlinico San Martino), Jürgen Ruland(Technical University of Munich), Armin Saalmüller(University of Veterinary Medicine Vienna), Yvan Saeys(Ghent University), Takashi Saito(RIKEN Center for Integrative Medical Sciences), Shimon Sakaguchi(The University of Osaka), Francisco Sala‐de‐Oyanguren(Ludwig Cancer Research), Yvonne Samstag(Heidelberg University), Sharon Sanderson(University of Oxford), Inga Sandrock(Medizinische Hochschule Hannover), Angela Santoni(Istituto Neurologico Mediterraneo), R. Bellmàs Sanz(Medizinische Hochschule Hannover), Marina Saresella(Don Carlo Gnocchi Foundation), Catherine Sautès‐Fridman(Centre de Recherche des Cordeliers), Birgit Sawitzki(Berlin Institute of Health at Charité - Universitätsmedizin Berlin), Linda Schadt(University of Zurich), Alexander Scheffold(Christian-Albrechts-Universität zu Kiel), Hans Ulrich Scherer(Leiden University Medical Center), Matthias Schiemann(Technical University of Munich), Frank A. Schildberg(University Hospital Bonn), Esther Schimisky(Miltenyi Biotec (Germany)), Andreas Schlitzer(University of Bonn), Josephine Schlosser(Freie Universität Berlin), Stephan Schmid(University Hospital Regensburg), Steffen Schmitt(German Cancer Research Center), Kilian Schober(Technical University of Munich), Daniel Schraivogel(European Molecular Biology Laboratory), Wolfgang Schuh(Friedrich-Alexander-Universität Erlangen-Nürnberg), Thomas Schüler(Otto-von-Guericke-Universität Magdeburg), Reiner Schulte(University of Cambridge), Axel Schulz(Leibniz Association), Sebastian Schulz(Friedrich-Alexander-Universität Erlangen-Nürnberg), Cristiano Scottà(King's College London), Daniel Scott‐Algara(Institut Pasteur), David P. Sester(Translational Research Institute), T. Vincent Shankey(Purdue University West Lafayette), Bruno Silva‐Santos(University of Lisbon), Anna Katharina Simon(University of Oxford), Katarzyna Sitnik(Helmholtz Centre for Infection Research), Silvano Sozzani(University of Brescia), Daniel E. Speiser(University of Lausanne), Josef Špidlen(Ashland (United States)), Anders Ståhlberg(University of Gothenburg), Alan M. Stall(BD Biosciences (United States)), Natalie Stanley(Stanford Medicine), Regina Stark(Sanquin), Christina Stehle(Leibniz Association), Tobit Steinmetz(Friedrich-Alexander-Universität Erlangen-Nürnberg), Hannes Stockinger(Medical University of Vienna), Yousuke Takahama(National Institutes of Health), Kiyoshi Takeda(The University of Osaka), Leonard Tan(Agency for Science, Technology and Research), Attila Tárnok(Fraunhofer Institute for Cell Therapy and Immunology), Gisa Tiegs(Universität Hamburg), Gergely Toldi(University of Szeged), Julia Tornack(Leibniz Association), Elisabetta Traggiai(Novartis (Switzerland)), Mohamed Trebak(Pennsylvania State University), Timothy Tree(National Health Service), Joe Trotter(BD Biosciences (United States)), John Trowsdale(University of Cambridge), Maria Tsoumakidou(Alexander Fleming Biomedical Sciences Research Center), Henning Ulrich(Universidade de São Paulo), Sophia Urbanczyk(Friedrich-Alexander-Universität Erlangen-Nürnberg), Willem van de Veen(University of Zurich), Maries van den Broek(University of Zurich), Edwin van der Pol(Amsterdam University Medical Centers), Sofie Van Gassen(Ghent University), Gert Van Isterdael(VIB-UGent Center for Inflammation Research), René A. W. van Lier(Sanquin), Marc Veldhoen(University of Lisbon), Salvador Vento‐Asturias(University of Bonn), Paulo Vieira(Institut Pasteur), David Voehringer(Friedrich-Alexander-Universität Erlangen-Nürnberg), Hans‐Dieter Volk(Berlin Institute of Health at Charité - Universitätsmedizin Berlin), Anouk von Borstel(Australian Research Council), Konrad von Volkmann, Ari Waisman(Johannes Gutenberg University Mainz), Rachael Walker(Babraham Institute), Paul K. Wallace(Roswell Park Comprehensive Cancer Center), Sa A. Wang(The University of Texas MD Anderson Cancer Center), Xin Wang(Westmead Institute), Michael D. Ward(Thermo Fisher Scientific (United States)), Kirsten A. Ward‐Hartstonge(University of British Columbia), Klaus Warnatz(University of Freiburg), Gary Warnes(University of London), Sarah Warth(Berlin-Brandenburger Centrum für Regenerative Therapien), Claudia Waskow(Leibniz Institute on Aging - Fritz Lipmann Institute (FLI)), James Watson(Middlesex University), Carsten Watzl(TU Dortmund University), Leonie Wegener(Miltenyi Biotec (Germany)), Thomas Weisenburger(Friedrich-Alexander-Universität Erlangen-Nürnberg), Annika Wiedemann(Leibniz Association), Jürgen Wienands(Universitätsmedizin Göttingen), Anneke Wilharm(Medizinische Hochschule Hannover), Robert J. Wilkinson(University of Cape Town), Gerald Willimsky(German Cancer Research Center), James B. Wing(The University of Osaka), Rieke Winkelmann(Christian-Albrechts-Universität zu Kiel), Thomas Winkler(Friedrich-Alexander-Universität Erlangen-Nürnberg), Oliver F. Wirz(University of Zurich), Alicia Y. W. Wong(Agency for Science, Technology and Research), Peter Wurst(University of Bonn), Jennie H. M. Yang(National Health Service), Juhao Yang(King's College London), Maria Yazdanbakhsh(Leiden University Medical Center), Liping Yu(BD Biosciences (United States)), Alice Yue(Simon Fraser University), Hanlin Zhang(University of Oxford), Yi Zhao(Sichuan University), Susanne Ziegler(Leibniz Institute of Virology (LIV)), Christina E. Zielinski(German Center for Infection Research), Jakob Zimmermann(University of Bern), Arturo Zychlinsky(Max Planck Institute for Infection Biology)
Cited by 986Open Access
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.
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