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Eur J Immunol. 2019 Oct;49(10):1457-1973. doi: 10.1002/eji.201970107.

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition).

Cossarizza A1, Chang HD2, Radbruch A2, Acs A3, Adam D4, Adam-Klages S5, Agace WW6,7, Aghaeepour N8, Akdis M9, Allez M10, Almeida LN11, Alvisi G12, Anderson G13, Andrä I14, Annunziato F15, Anselmo A16, Bacher P4,17, Baldari CT18, Bari S19,20, Barnaba V21,22,23, Barros-Martins J24, Battistini L25, Bauer W26, Baumgart S2, Baumgarth N27, Baumjohann D28, Baying B29, Bebawy M30, Becher B31,32, Beisker W33, Benes V29, Beyaert R34, Blanco A35, Boardman DA36,37, Bogdan C38,39, Borger JG40, Borsellino G41, Boulais PE42,43, Bradford JA44, Brenner D45,46,47, Brinkman RR48,49, Brooks AES50, Busch DH14,51,52, Büscher M53, Bushnell TP54, Calzetti F55, Cameron G56, Cammarata I21, Cao X57, Cardell SL58, Casola S59, Cassatella MA55, Cavani A60, Celada A61, Chatenoud L62, Chattopadhyay PK63, Chow S64, Christakou E65,66, Čičin-Šain L67, Clerici M68,69,70, Colombo FS16, Cook L37,71, Cooke A72, Cooper AM73, Corbett AJ56, Cosma A74, Cosmi L15, Coulie PG75, Cumano A76, Cvetkovic L77, Dang VD2, Dang-Heine C78, Davey MS79,80, Davies D81, De Biasi S82, Del Zotto G83, Dela Cruz GV84, Delacher M85,86, Della Bella S87, Dellabona P88, Deniz G89, Dessing M90, Di Santo JP91,92, Diefenbach A2,93,94, Dieli F95, Dolf A96, Dörner T2,97, Dress RJ98, Dudziak D99, Dustin M100, Dutertre CA101,98, Ebner F102, Eckle SBG56, Edinger M85,103, Eede P104, Ehrhardt GRA105, Eich M106, Engel P107, Engelhardt B108, Erdei A109, Esser C110, Everts B111, Evrard M98, Falk CS112, Fehniger TA113, Felipo-Benavent M114, Ferry H115, Feuerer M85,86, Filby A116, Filkor K117, Fillatreau S118, Follo M119,120, Förster I121, Foster J122, Foulds GA123, Frehse B11, Frenette PS42,43,124, Frischbutter S2,125, Fritzsche W126, Galbraith DW127,128, Gangaev A129, Garbi N130, Gaudilliere B131, Gazzinelli RT132,133, Geginat J134, Gerner W135,136, Gherardin NA56, Ghoreschi K137, Gibellini L82, Ginhoux F98,138,139, Goda K140,141,142, Godfrey DI56, Goettlinger C143, González-Navajas JM144,145, Goodyear CS146, Gori A147, Grogan JL148, Grummitt D122, Grützkau A2, Haftmann C31, Hahn J149, Hammad H150, Hämmerling G151, Hansmann L94,152,153, Hansson G154, Harpur CM155, Hartmann S102, Hauser A103, Hauser AE2,156,157, Haviland DL158, Hedley D64, Hernández DC2,159, Herrera G160, Herrmann M149, Hess C161,162, Höfer T163, Hoffmann P85,103, Hogquist K164, Holland T130, Höllt T165,166, Holmdahl R167, Hombrink P168,169, Houston JP170, Hoyer BF171, Huang B172, Huang FP173, Huber JE28, Huehn J174, Hundemer M175, Hunter CA176, Hwang WYK177,20,178, Iannone A179, Ingelfinger F31, Ivison SM36,37, Jäck HM77, Jani PK2,180, Jávega B181, Jonjic S182, Kaiser T2, Kalina T183, Kamradt T184, Kaufmann SHE180, Keller B185,186, Ketelaars SLC129, Khalilnezhad A98,187, Khan S105, Kisielow J188, Klenerman P115, Knopf J149, Koay HF56, Kobow K189, Kolls JK190, Kong WT98, Kopf M188, Korn T191, Kriegsmann K175, Kristyanto H192, Kroneis T193, Krueger A194, Kühne J112, Kukat C195, Kunkel D196,197, Kunze-Schumacher H194, Kurosaki T198, Kurts C130, Kvistborg P129, Kwok I98,199, Landry J29, Lantz O200, Lanuti P201, LaRosa F68,70, Lehuen A202, LeibundGut-Landmann S203, Leipold MD204, Leung LYT105, Levings MK36,37,205, Lino AC2,97, Liotta F15, Litwin V206, Liu Y105, Ljunggren HG207, Lohoff M208, Lombardi G209, Lopez L210, López-Botet M211, Lovett-Racke AE212, Lubberts E213, Luche H214, Ludewig B215, Lugli E12,16, Lunemann S216, Maecker HT217, Maggi L15, Maguire O218, Mair F219, Mair KH135,136, Mantovani A220,221, Manz RA11, Marshall AJ222, Martínez-Romero A223, Martrus G216, Marventano I68,70, Maslinski W224, Matarese G225, Mattioli AV82,226, Maueröder C227,228, Mazzoni A15, McCluskey J56, McGrath M2, McGuire HM229, McInnes IB146, Mei HE2, Melchers F2,180, Melzer S230, Mielenz D77, Miller SD231, Mills KHG232, Minderman H218, Mjösberg J207,233, Moore J234, Moran B232, Moretta L235, Mosmann TR236, Müller S237, Multhoff G238,239, Muñoz LE149, Münz C31,32, Nakayama T240, Nasi M82, Neumann K241, Ng LG98,187,199,242,243, Niedobitek A2, Nourshargh S244, Núñez G245, O'Connor JE181, Ochel A241, Oja A169, Ordonez D246, Orfao A247, Orlowski-Oliver E248, Ouyang W249, Oxenius A250, Palankar R251, Panse I2, Pattanapanyasat K252, Paulsen M246, Pavlinic D29, Penter L153, Peterson P253, Peth C53, Petriz J254, Piancone F68,70, Pickl WF255, Piconese S21,23, Pinti M256, Pockley AG123,257, Podolska MJ149,258, Poon Z177, Pracht K77, Prinz I24, Pucillo CEM259, Quataert SA236, Quatrini L235, Quinn KM260,261, Radbruch H104, Radstake TRDJ262, Rahmig S263, Rahn HP264, Rajwa B265, Ravichandran G241, Raz Y266, Rebhahn JA236, Recktenwald D267, Reimer D77, Reis E Sousa C268, Remmerswaal EBM168,269, Richter L270, Rico LG254, Riddell A81, Rieger AM271, Robinson JP272, Romagnani C2,159, Rubartelli A273, Ruland J274, Saalmüller A135, Saeys Y275,276, Saito T277, Sakaguchi S198, Sala-de-Oyanguren F278, Samstag Y279, Sanderson S280, Sandrock I24, Santoni A281, Sanz RB112, Saresella M68,70, Sautes-Fridman C282, Sawitzki B283, Schadt L31,32, Scheffold A4, Scherer HU192, Schiemann M14, Schildberg FA284, Schimisky E285, Schlitzer A286, Schlosser J102, Schmid S287, Schmitt S288, Schober K14, Schraivogel D289, Schuh W77, Schüler T290, Schulte R291, Schulz AR2, Schulz SR77, Scottá C209, Scott-Algara D292, Sester DP293, Shankey TV294, Silva-Santos B295, Simon AK100, Sitnik KM67, Sozzani S296, Speiser DE297, Spidlen J298, Stahlberg A299, Stall AM300, Stanley N8, Stark R168,169, Stehle C2,159, Steinmetz T77, Stockinger H301, Takahama Y302, Takeda K198, Tan L98,187, Tárnok A303,304,305, Tiegs G241, Toldi G117, Tornack J2,306, Traggiai E307, Trebak M308, Tree TIM65,66, Trotter J300, Trowsdale J72, Tsoumakidou M309, Ulrich H310, Urbanczyk S77, van de Veen W9,311, van den Broek M31,32, van der Pol E312, Van Gassen S275,276, Van Isterdael G313, van Lier RAW169, Veldhoen M295, Vento-Asturias S130, Vieira P76, Voehringer D314, Volk HD315, von Borstel A79,80, von Volkmann K316, Waisman A317, Walker RV318, Wallace PK319, Wang SA320, Wang XM321, Ward MD44, Ward-Hartstonge KA36, Warnatz K185,186, Warnes G322, Warth S197, Waskow C263,323, Watson JV324, Watzl C325, Wegener L53, Weisenburger T3, Wiedemann A2,97, Wienands J326, Wilharm A24, Wilkinson RJ327,328,329, Willimsky G330, Wing JB198, Winkelmann R4, Winkler TH3, Wirz OF9, Wong A98, Wurst P331, Yang JHM65,66, Yang J174, Yazdanbakhsh M111, Yu L332, Yue A333, Zhang H100, Zhao Y334, Ziegler SM216, Zielinski C51,335,336, Zimmermann J337, Zychlinsky A180.

Author information

1
Department of Medical and Surgical Sciences for Children and Adults, Univ. of Modena and Reggio Emilia School of Medicine, Modena, Italy.
2
Deutsches Rheuma-Forschungszentrum (DRFZ), an Institute of the Leibniz Association, Berlin, Germany.
3
Department of Biology, Nikolaus-Fiebiger-Center for Molecular Medicine, Friedrich-Alexander-University Erlangen-Nuremberg, Erlangen, Germany.
4
Institut für Immunologie, Christian-Albrechts-Universität zu Kiel, Kiel, Germany.
5
Institut für Transfusionsmedizin, Universitätsklinik Schleswig-Holstein, Kiel, Germany.
6
Mucosal Immunology group, Department of Health Technology, Technical University of Denmark, Kgs. Lyngby, Denmark.
7
Immunology Section, Lund University, Lund, Sweden.
8
Departments of Anesthesiology, Pain and Perioperative Medicine, Biomedical Data Sciences; and Pediatrics, Stanford University, Stanford, CA, USA.
9
Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Davos, Switzerland.
10
Université de Paris, Institut de Recherche Saint-Louis, INSERM U1160, and Gastroenterology Department, Hôpital Saint-Louis - APHP, Paris, France.
11
Institute for Systemic Inflammation Research, University of Luebeck, Luebeck, Germany.
12
Laboratory of Translational Immunology, Humanitas Clinical and Research Center, Rozzano, Italy.
13
University of Birmingham, Birmingham, UK.
14
Institut für Medizinische Mikrobiologie, Immunologie und Hygiene, Technische Universität München, Munich, Germany.
15
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
16
Flow Cytometry Core, Humanitas Clinical and Research Center, Milan, Italy.
17
Institut für Klinische Molekularbiologie, Christian-Albrechts Universität zu Kiel, Germany.
18
Department of Life Sciences, University of Siena, Siena, Italy.
19
Division of Medical Sciences, National Cancer Centre Singapore, Singapore.
20
Cancer & Stem Cell Biology, Duke-NUS Medical School, Singapore.
21
Dipartimento di Medicina Interna e Specialità Mediche, Sapienza Università di Roma, Rome, Italy.
22
Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Rome, Italy.
23
Istituto Pasteur - Fondazione Cenci Bolognetti, Rome, Italy.
24
Institute of Immunology, Hannover Medical School, Hannover, Germany.
25
IRCCS Fondazione Santa Lucia, Rome, Italy.
26
Division of Immunology, Allergy and Infectious Diseases, Department of Dermatology, Medical University of Vienna, Vienna, Austria.
27
Center for Comparative Medicine & Dept. Pathology, Microbiology & Immunology, University of California, Davis, CA, USA.
28
Institute for Immunology, Faculty of Medicine, Biomedical Center, LMU Munich, Planegg-Martinsried, Germany.
29
Genomics Core Facility, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
30
Discipline of Pharmacy, Graduate School of Health, The University of Technology Sydney, Sydney, NSW, Australia.
31
Institute of Experimental Immunology, University of Zurich, Zurich, Switzerland.
32
Comprehensive Cancer Center Zurich, Switzerland.
33
Flow Cytometry Laboratory, Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München, German Research Center for Environmental Health, München, Germany.
34
Department of Biomedical Molecular Biology, Center for Inflammation Research, Ghent University - VIB, Ghent, Belgium.
35
Flow Cytometry Core Technologies, UCD Conway Institute, University College Dublin, Dublin, Ireland.
36
Department of Surgery, The University of British Columbia, Vancouver, Canada.
37
BC Children's Hospital Research Institute, Vancouver, Canada.
38
Mikrobiologisches Institut - Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen, Erlangen, Germany.
39
Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg and Medical Immunology Campus Erlangen, Erlangen, Germany.
40
Department of Immunology and Pathology, Monash University, Melbourne, Victoria, Australia.
41
Neuroimmunology and Flow Cytometry Units, Fondazione Santa Lucia IRCCS, Rome, Italy.
42
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY, USA.
43
The Ruth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Bronx, New York, USA.
44
Thermo Fisher Scientific, Eugene, Oregon, USA.
45
Luxembourg Institute of Health, Department of Infection and Immunity, Experimental and Molecular Immunology, Esch-sur-Alzette, Luxembourg.
46
Odense University Hospital, Odense Research Center for Anaphylaxis, University of Southern Denmark, Department of Dermatology and Allergy Center, Odense, Denmark.
47
Luxembourg Centre for Systems Biomedicine (LCSB), University of Luxembourg, Belvaux, Luxembourg.
48
Department of Medical Genetics, University of British Columbia, Vancouver, BC, Canada.
49
Terry Fox Laboratory, BC Cancer, Vancouver, BC, Canada.
50
University of Auckland, School of Biological Sciences, Maurice Wilkins Center, Auckland, New Zealand.
51
German Center for Infection Research (DZIF), Munich, Germany.
52
Focus Group ''Clinical Cell Processing and Purification", Institute for Advanced Study, Technische Universität München, Munich, Germany.
53
Biophysics, R&D Engineering, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
54
Department of Pediatrics and Shared Resource Laboratories, University of Rochester Medical Center, Rochester, NY, USA.
55
University of Verona, Department of Medicine, Section of General Pathology, Verona, Italy.
56
Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria, Australia.
57
National Key Laboratory of Medical Immunology, Nankai University, Tianjin, China.
58
Department of Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden.
59
The FIRC Institute of Molecular Oncology (FOM), Milan, Italy.
60
National Institute for Health, Migration and Poverty (INMP), Rome, Italy.
61
Macrophage Biology Group, School of Biology, University of Barcelona, Barcelona, Spain.
62
Université Paris Descartes, Institut National de la Santé et de la Recherche Médicale, Paris, France.
63
Isaac and Laura Perlmutter Cancer Center, NYU-Langone Medical Center, New York, USA.
64
Divsion of Medical Oncology and Hematology, Princess Margaret Hospital, Toronto, Ontario, Canada.
65
Department of Immunobiology, School of Immunology and Microbial Sciences, King's College London, UK.
66
National Institutes of Health Research Biomedical Research Centre at Guy's and St. Thomas' National Health Service, Foundation Trust and King's College London, UK.
67
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
68
IRCCS Fondazione Don Carlo Gnocchi, Milan, Italy.
69
Department of Physiopathology and Transplants, University of Milan, Milan, Italy.
70
Milan Center for Neuroscience, University of Milano-Bicocca, Milan, Italy.
71
Department of Medicine, The University of British Columbia, Vancouver, Canada.
72
Department of Pathology, University of Cambridge, Cambridge, UK.
73
Department of Respiratory Sciences, University of Leicester, Leicester, UK.
74
National Cytometry Platform, Luxembourg Institute of Health, Department of Infection and Immunity, Esch-sur-Alzette, Luxembourg.
75
de Duve Institute, Université catholique de Louvain, Brussels, Belgium.
76
Unit Lymphopoiesis, Department of Immunology, Institut Pasteur, Paris, France.
77
Division of Molecular Immunology, Nikolaus-Fiebiger-Center, Dept. of Internal Medicine III, University of Erlangen-Nuremberg, Erlangen, Germany.
78
Clinical Research Unit, Berlin Institute of Health (BIH), Charite Universitätsmedizin Berlin, Berlin, Germany.
79
Infection and Immunity Program and Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia.
80
Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Monash University, Clayton, Victoria, Australia.
81
Flow Cytometry Scientific Technology Platform, The Francis Crick Institute, London, UK.
82
Department of Surgery, Medicine, Dentistry and Morphological Sciences, Univ. of Modena and Reggio Emilia, Modena, Italy.
83
IRCCS Istituto Giannina Gaslini, Genova, Italy.
84
Novo Nordisk Foundation Center for Stem Cell Biology - DanStem, University of Copenhagen, Copenhagen, Denmark.
85
Regensburg Center for Interventional Immunology (RCI), Regensburg, Germany.
86
Chair for Immunology, University Regensburg, Germany.
87
Department of Medical Biotechnologies and Translational Medicine, University of Milan, Milan, Italy.
88
Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy.
89
Istanbul University, Aziz Sancar Institute of Experimental Medicine, Department of Immunology, Istanbul, Turkey.
90
Sony Europe Ltd, Weybridge, UK.
91
Innate Immunty Unit, Department of Immunology, Institut Pasteur, Paris, France.
92
Institut Pasteur, Inserm U1223, Paris, France.
93
Charité - Universitätsmedizin Berlin, Laboratory of Innate Immunity, Department of Microbiology, Infectious Diseases and Immunology, Berlin, Germany.
94
Berlin Institute of Health (BIH), Berlin, Germany.
95
University of Palermo, Central Laboratory of Advanced Diagnosis and Biomedical Research, Department of Biomedicine, Neurosciences and Advanced Diagnostics, Palermo, Italy.
96
Flow Cytometry Core Facility, Institute of Experimental Immunology, University of Bonn, Bonn, Germany.
97
Dept. Medicine/Rheumatology and Clinical Immunology, Charité Universitätsmedizin Berlin, Germany.
98
Singapore Immunology Network (SIgN), A*STAR (Agency for Science, Technology and Research), Biopolis, Singapore.
99
Department of Dermatology, Laboratory of Dendritic Cell Biology, Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), University Hospital Erlangen, Erlangen, Germany.
100
Kennedy Institute of Rheumatology, University of Oxford, Oxford, UK.
101
Program in Emerging Infectious Disease, Duke-NUS Medical School, Singapore.
102
Institute of Immunology, Centre for Infection Medicine, Department of Veterinary Medicine, Freie Universität Berlin, Germany.
103
Department of Internal Medicine III, University Hospital Regensburg, Germany.
104
Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Department of Neuropathology, Germany.
105
Department of Immunology, University of Toronto, Toronto, ON, Canada.
106
Heidelberg Institute for Stem Cell Technology and Experimental Medicine (HI-STEM gGmbH), Heidelberg, Germany.
107
University of Barcelona, Faculty of Medicine and Health Sciences, Department of Biomedical Sciences, Barcelona, Spain.
108
Theodor Kocher Institute, University of Bern, Bern, Switzerland.
109
Department of Immunology, University L. Eotvos, Budapest, Hungary.
110
Leibniz Research Institute for Environmental Medicine, Düsseldorf, Germany.
111
Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands.
112
Institute of Transplant Immunology, Hannover Medical School, MHH, Hannover, Germany.
113
Division of Oncology, Washington University School of Medicine, St. Louis, MO, USA.
114
Laboratory of Cytomics, Joint Research Unit CIPF-UVEG, Principe Felipe Research Center, Valencia, Spain.
115
Experimental Medicine Division, Nuffield Department of Medicine, University of Oxford, Oxford, UK.
116
The Flow Cytometry Core Facility, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK.
117
MDQuest Ltd., Szeged, Hungary.
118
Institut Necker-Enfants Malades, Université Paris Descartes Sorbonne Paris Cité, Faculté de Médecine, AP-HP, Hôpital Necker Enfants Malades, INSERM U1151-CNRS UMR 8253, Paris, France.
119
Department of Medicine I, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
120
Universitaetsklinikum Freiburg, Lighthouse Core Facility, Zentrum für Translationale Zellforschung, Klinik für Innere Medizin I, Freiburg, Germany.
121
Immunology and Environment, LIMES Institute, University of Bonn, Bonn, Germany.
122
Owl Biomedical Inc., Santa Barbara, USA.
123
John van Geest Cancer Research Centre, Nottingham Trent University, Nottingham, UK.
124
Department of Medicine, Albert Einstein College of Medicine, Bronx, NY, USA.
125
Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Department of Dermatology, Venereology and Allergology.
126
Nanobiophotonics Department, Leibniz Institute of Photonic Technology (IPHT), Jena, Germany.
127
School of Plant Sciences and Bio5 Institute, University of Arizona, Tucson, USA.
128
Honorary Dean of Life Sciences, Henan University, Kaifeng, China.
129
Division of Molecular Oncology and Immunology, the Netherlands Cancer Institute, Amsterdam, The Netherlands.
130
Institute of Experimental Immunology, University of Bonn, Germany.
131
Stanford Department of Anesthesiology, Perioperative and Pain Medicine, Stanford University School of Medicine, CA, USA.
132
Fundação Oswaldo Cruz - Minas, Laboratory of Immunopatology, Belo Horizonte, MG, Brazil.
133
Department of Mecicine, University of Massachusetts Medical School, Worcester, MA, USA.
134
INGM - Fondazione Istituto Nazionale di Genetica Molecolare "Ronmeo ed Enrica Invernizzi", Milan, Italy.
135
Institute of Immunology, Department of Pathobiology, University of Veterinary Medicine Vienna, Austria.
136
Christian Doppler Laboratory for Optimized Prediction of Vaccination Success in Pigs, Institute of Immunology, Department of Pathobiology, University of Veterinary Medicine Vienna, Austria.
137
Department of Dermatology, Venereology and Allergology, Charité - Universitätsmedizin Berlin, Berlin, Germany.
138
Translational Immunology Institute, SingHealth Duke-NUS Academic Medical Centre, Singapore.
139
Shanghai Institute of Immunology, Department of Immunology and Microbiology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
140
Department of Bioengineering, University of California, Los Angeles, California, USA.
141
Department of Chemistry, University of Tokyo, Tokyo, Japan.
142
Institute of Technological Sciences, Wuhan University, Wuhan, China.
143
Institut für Genetik, Universität zu Köln, Köln, Germany.
144
Alicante Institute for Health and Biomedical Research (ISABIAL), Alicante, Spain.
145
Networked Biomedical Research Center for Hepatic and Digestive Diseases (CIBERehd), Madrid, Spain.
146
Institute of Infection Immunity and Inflammation, College of Medical Veterinary and Life Sciences, University of Glasgow, Glasgow Biomedical Research Centre, Glasgow, UK.
147
Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico, University of Milan.
148
Cancer Immunology Research, Genentech, South San Francisco, CA, USA.
149
Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Department of Medicine 3, Rheumatology and Immunology, Universitätsklinikum Erlangen, Erlangen.
150
Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Zwijnaarde, Belgium.
151
German Cancer Research Center (DKFZ), Heidelberg, Germany.
152
German Cancer Consortium (DKTK), partner site Berlin, Berlin, Germany.
153
Department of Hematology, Oncology, and Tumor Immunology, Charité - Universitätsmedizin Berlin, Campus Virchow Klinikum, Berlin, Germany.
154
Department of Medicine and Center for Molecular Medicine at Karolinska University Hospital, Solna, Sweden.
155
Murdoch Children's Research Institute, Parkville, Victoria, Australia.
156
Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin.
157
Department of Rheumatology and Clinical Immunology, Berlin Institute of Health, Berlin, Germany.
158
Flow Cytometry, Houston Methodist Hospital Research Institute, Houston, TX, USA.
159
Charité - Universitätsmedizin Berlin, Medical Department I, Division of Gastroenterology, Infectiology and Rheumatology, Berlin, Germany.
160
Cytometry Service, Incliva Foundation. Clinic Hospital and Faculty of Medicine, University of Valencia, Valencia, Spain.
161
Immunobiology Laboratory, Department of Biomedicine, University and University Hospital Basel, Basel, Switzerland.
162
Cambridge Institute of Therapeutic Immunology & Infectious Disease, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK.
163
German Cancer Research Center (DKFZ), Division of Theoretical Systems Biology, Heidelberg, Germany.
164
Center for Immunology, University of Minnesota, Minneapolis, MN, USA.
165
Leiden Computational Biology Center, Leiden University Medical Center, Leiden, The Netherlands.
166
Computer Graphics and Visualization, Department of Intelligent Systems, TU Delft, Delft, The Netherlands.
167
Karolinska Institutet Biomedicum, Solna, Sweden.
168
Department of Experimental Immunology, Amsterdam Infection and Immunity Institute, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.
169
Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.
170
Department of Chemical & Materials Engineering, New Mexico State University, Las Cruces, NM, USA.
171
Rheumatologie/Klinische Immunologie, Klinik für Innere Medizin I und Exzellenzzentrum Entzündungsmedizin, Universitätsklinikum Schleswig-Holstein, Kiel, Germany.
172
Department of Immunology & National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences (CAMS) & Peking Union Medical College, Beijing, China.
173
Institute for Advanced Study (IAS), Shenzhen University, Shenzhen, China.
174
Experimental Immunology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
175
Department of Hematology, Oncology and Rheumatology, University Heidelberg, Heidelberg, Germany.
176
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.
177
Department of Hematology, Singapore General Hospital, Singapore.
178
Executive Offices, National Cancer Centre Singapore, Singapore.
179
Department of Diagnostic Medicine, Clinical and Public Health, Univ. of Modena and Reggio Emilia, Modena, Italy.
180
Max Planck Institute for Infection Biology, Berlin, Germany.
181
Laboratory of Cytomics, Joint Research Unit CIPF-UVEG, Department of Biochemistry and Molecular Biology, University of Valencia, Valencia, Spain.
182
Department of Histology and Embryology/Center for Proteomics, Faculty of Medicine, University of Rijeka, Rijeka, Croatia.
183
Department of Paediatric Haematology and Oncology, Second Faculty of Medicine, Charles University, Prague, Czech Republic.
184
Jena University Hospital, Institute of Immunology, Jena, Germany.
185
Department of Rheumatology and Clinical Immunology, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
186
Center for Chronic Immunodeficiency, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
187
Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
188
Institute of Molecular Health Sciences, ETH Zurich, Zürich, Switzerland.
189
Department of Neuropathology, Universitätsklinikum Erlangen, Germany.
190
John W Deming Endowed Chair in Internal Medicine, Center for Translational Research in Infection and Inflammation Tulane School of Medicine, New Orleans, LA, USA.
191
Department of Neurology, Technical University of Munich, Munich, Germany.
192
Department of Rheumatology, Leiden University Medical Center, Leiden, The Netherlands.
193
Division of Cell Biology, Histology & Embryology, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria.
194
Institute for Molecular Medicine, Goethe University Frankfurt, Frankfurt am Main, Germany.
195
FACS & Imaging Core Facility, Max Planck Institute for Biology of Ageing, Cologne, Germany.
196
Flow & Mass Cytometry Core Facility, Charité - Universitätsmedizin Berlin and Berlin Institute of Health, Berlin, Germany.
197
BCRT Flow Cytometry Lab, Berlin-Brandenburg Center for Regenerative Therapies, Charité - Universitätsmedizin Berlin.
198
WPI Immunology Frontier Research Center, Osaka University, Osaka, Japan.
199
School of Biological Sciences, Nanyang Technological University, Singapore.
200
INSERM U932, PSL University, Institut Curie, Paris, France.
201
Department of Medicine and Aging Sciences, Centre on Aging Sciences and Translational Medicine (Ce.S.I.-Me.T.), University "G. d'Annunzio" of Chieti-Pescara, Chieti, Italy.
202
Institut Cochin, CNRS8104, INSERM1016, Department of Endocrinology, Metabolism and Diabetes, Université de Paris, Paris, France.
203
Section of Immunology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland.
204
The Human Immune Monitoring Center (HIMC), Institute for Immunity, Transplantation and Infection, Stanford University School of Medicine, CA, USA.
205
School of Biomedical Engineering, The University of British Columbia, Vancouver, Canada.
206
Caprion Biosciences, ImmuneCarta, Montréal, Quebec, Canada.
207
Center for Infectious Medicine, Department of Medicine Huddinge, ANA Futura, Karolinska Institutet, Stockholm, Sweden.
208
Inst. f. Med. Mikrobiology and Hospital Hygiene, University of Marburg, Germany.
209
King's College London, "Peter Gorer" Department of Immunobiology, London, UK.
210
Beckman Coulter, Inc, Miami, FL, USA.
211
IMIM(Hospital de Mar Medical Research Institute), University Pompeu Fabra, Barcelona, Spain.
212
Department of Microbial Infection and Immunity, Ohio State University, Columbus, OH, USA.
213
Department of Rheumatology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.
214
Centre d'Immunophénomique - CIPHE (PHENOMIN), Aix Marseille Université (UMS3367), Inserm (US012), CNRS (UMS3367), Marseille, France.
215
Institute of Immunobiology, Kantonsspital St.Gallen, St. Gallen, Switzerland.
216
Department of Virus Immunology, Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.
217
Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Stanford, CA, USA.
218
Flow and Image Cytometry Shared Resource, Roswell Park Comprehensive Cancer Center, Buffalo, NY, USA.
219
Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, USA.
220
Istituto Clinico Humanitas IRCCS and Humanitas University, Pieve Emanuele, Milan, Italy.
221
William Harvey Research Institute, Queen Mary University, London, United Kingdom.
222
Department of Immunology, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada.
223
Cytomics Technological Service, Principe Felipe Research Center, Valencia, Spain.
224
National Institute of Geriatrics, Rheumatology and Rehabilitation, Department of Pathophysiology and Immunology, Warsaw, Poland.
225
Treg Cell Lab, Dipartimento di Medicina Molecolare e Biotecologie Mediche, Università di Napoli Federico II and Istituto per l'Endocrinologia e l'Oncologia Sperimentale, Consiglio Nazionale delle Ricerche (IEOS-CNR), Napoli, Italy.
226
Lab of Clinical and Experimental Immunology, Humanitas Clinical and Research Center, Rozzano, Milan, Italy.
227
Cell Clearance in Health and Disease Lab, VIB Center for Inflammation Research, Ghent, Belgium.
228
Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
229
Ramaciotti Facility for Human Systems Biology, and Discipline of Pathology, The University of Sydney, Camperdown, Australia.
230
Clinical Trial Center Leipzig, University Leipzig, Leipzig, Germany.
231
Interdepartmental Immunobiology Center, Dept. of Microbiology-Immunology, Northwestern Univ. Medical School, Chicago, IL, USA.
232
Trinity College Dublin, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Dublin, Ireland.
233
Department of Clinical and Experimental Medine, Linköping University, Linköping, Sweden.
234
Abramson Cancer Center Flow Cytometry and Cell Sorting Shared Resource, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA.
235
Department of Immunology, IRCCS Bambino Gesu Children's Hospital, Rome, Italy.
236
David H. Smith Center for Vaccine Biology and Immunology, University of Rochester Medical Center, Rochester, NY, USA.
237
Centre for Environmental Research - UFZ, Department Environmental Microbiology, Leipzig, Germany.
238
Institute for Innovative Radiotherapy (iRT), Experimental Immune Biology, Helmholtz Zentrum München, Neuherberg, Germany.
239
Radiation Immuno-Oncology Group, Center for Translational Cancer Research Technische Universität München (TranslaTUM), Klinikum rechts der Isar, Munich, Germany.
240
Department of Immunology, Graduate School of Medicine, Chiba University, Chiba city, Chiba, Japan.
241
Institute of Experimental Immunology and Hepatology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
242
Discipline of Dermatology, University of Sydney, Sydney, New South Wales, Australia.
243
State Key Laboratory of Experimental Hematology, Institute of Hematology, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China.
244
Barts and The London School of Medicine and Dentistry, Queen Mary University of London, UK.
245
Department of Pathology and Rogel Cancer Center, the University of Michigan, Ann Arbor, Michigan, USA.
246
Flow Cytometry Core Facility, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
247
Department of Medicine, Cancer Research Centre (IBMCC-CSIC/USAL), Cytometry Service, University of Salamanca, CIBERONC and Institute for Biomedical Research of Salamanca (IBSAL), Salamanca, Spain.
248
Burnet Institute, AMREP Flow Cytometry Core Facility, Melbourne, Victoria, Australia.
249
Inflammation and Oncology, Research, Amgen Inc, South San Francisco, USA.
250
ETH Zurich, Institute of Microbiology, Zurich, Switzerland.
251
Department of Transfusion Medicine, Institute of Immunology and Transfusion Medicine, University Medicine Greifswald, Greifswald, Germany.
252
Center of Excellence for Flow Cytometry, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
253
Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia.
254
Functional Cytomics Group, Josep Carreras Leukaemia Research Institute, Campus ICO-Germans Trias i Pujol, Universitat Autònoma de Barcelona, UAB, Badalona, Spain.
255
Institute of Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria.
256
Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
257
Chromocyte Limited, Electric Works, Sheffield, UK.
258
Department for Internal Medicine 3, Institute for Rheumatology and Immunology, AG Munoz, Universitätsklinikum Erlangen, Erlangen, Germany.
259
Department of Immunology, University of Udine, Udine, Italy.
260
School of Biomedical and Health Sciences, RMIT University, Bundoora, Victoria, Australia.
261
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
262
Department of Rheumatology and Clinical Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
263
Regeneration in Hematopoiesis, Leibniz-Institute on Aging, Fritz-Lipmann-Institute (FLI), Jena, Germany.
264
Preparative Flow Cytometry, Max-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.
265
Bindley Biosciences Center, Purdue University, West Lafayette, IN, USA.
266
Department of Internal Medicine, Groene Hart Hospital, Gouda, The Netherlands.
267
Desatoya LLC, Reno, NV, USA.
268
Immunobiology Laboratory, The Francis Crick Institute, London, UK.
269
Renal Transplant Unit, Division of Internal Medicine, Academic Medical Centre, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.
270
Core Facility Flow Cytometry, Biomedical Center, Ludwig-Maximilians-University Munich, Germany.
271
Department of Medical Microbiology and Immunology, University of Alberta, Alberta, Canada.
272
Purdue University Cytometry Laboratories, Purdue University, West Lafayette, IN, USA.
273
Cell Biology Unit, IRCCS Ospedale Policlinico San Martino, Genova, Italy.
274
Institut für Klinische Chemie und Pathobiochemie, Fakultät für Medizin, Technische Universität München, München, Germany.
275
Data Mining and Modeling for Biomedicine, VIB-UGent Center for Inflammation Research, Ghent, Belgium.
276
Department of Applied Mathematics, Computer Science and Statistics, Ghent University, Ghent, Belgium.
277
RIKEN Center for Integrative Medical Sciences, Yokohama, Japan.
278
Flow Cytometry Facility, Ludwig Cancer Institute, Faculty of Medicine and Biology, University of Lausanne, Epalinges, Switzerland.
279
Heidelberg University, Institute of Immunology, Section of Molecular Immunology, Heidelberg, Germany.
280
Translational Immunology Laboratory, NIHR BRC, University of Oxford, Kennedy Institute of Rheumatology, Oxford, UK.
281
Department of Molecular Medicine, Sapienza University of Rome, IRCCS, Neuromed, Pozzilli, Italy.
282
Centre de Recherches des Cordeliers, Equipe Cancer et Immunité anti-tumorale, Paris, France.
283
Charité - Universitätsmedizin Berlin, and Berlin Institute of Health, Institute of Medical Immunology, Berlin, Germany.
284
Clinic for Orthopedics and Trauma Surgery, University Hospital Bonn, Bonn, Germany.
285
R&D Reagents, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
286
Quantitative Systems Biology, Life & Medical Sciences Institute, University of Bonn, Bonn, Germany.
287
Internal Medicine I, University Hospital Regensburg, Germany.
288
Flow Cytometry Core Facility, German Cancer Research Centre (DKFZ), Heidelberg, Germany.
289
Genome Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
290
Institute of Molecular and Clinical Immunology, Otto-von-Guericke University, Magdeburg, Germany.
291
University of Cambridge, Cambridge Institute for Medical Research, Cambridge, UK.
292
Institut Pasteur, Cellular Lymphocytes Biology, Immunology Departement, Paris, France.
293
TRI Flow Cytometry Suite (TRI.fcs), Translational Research Institute, Wooloongabba, QLD, Australia.
294
AsedaSciences, West Lafayette, IN, USA.
295
Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, Portugal.
296
Dept. Molecular Translational Medicine, University of Brescia, Brescia, Italy.
297
Department of Oncology, University of Lausanne and CHUV, Epalinges, Switzerland.
298
BD Life Sciences, Ashland, OR, USA.
299
Lundberg Laboratory for Cancer, Department of Pathology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden.
300
BD Life Sciences, San Diego, CA, USA.
301
Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria.
302
National Institutes of Health, Bethesda, MD, USA.
303
Departement for Therapy Validation, Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany.
304
Institute for Medical Informatics, Statistics and Epidemiology (IMISE), University of Leipzig, Leipzig, Germany.
305
Department of Precision Instruments, Tsinghua University, Beijing, China.
306
BioGenes GmbH, Berlin, Germany.
307
Novartis Biologics Center, Mechanistic Immunology Unit, Novartis Institute for Biomedical Research, NIBR, Basel, Switzerland.
308
Department of Cellular and Molecular Physiology, Penn State University College of Medicine, PA, United States.
309
Institute of Bioinnovation, BSRC Alexander Fleming, Vari, Greece.
310
Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, SP, Brazil.
311
Christine Kühne Center for Allergy Research and Education (CK-CARE), Davos, Switzerland.
312
Vesicle Observation Center, Biomedical Engineering & Physics, Laboratory Experimental Clinical Chemistry, Amsterdam University Medical Centers, Location AMC, The Netherlands.
313
VIB Flow Core, VIB Center for Inflammation Research, Ghent, Belgium.
314
Department of Infection Biology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nuremberg (FAU), Erlangen, Germany.
315
BIH Center for Regenerative Therapies (BCRT) Charité Universitätsmedizin Berlin and Berlin Institute of Health, Core Unit ImmunoCheck.
316
A·P·E-Applied Physics & Electronics, Berlin, Germany.
317
Institute for Molecular Medicine, University Medical Center of the Johannes Gutenberg University of Mainz, Mainz, Germany.
318
Flow Cytometry Facility, Babraham Institute, Cambridge, UK.
319
Roswell Park Comprehensive Cancer Center, Elm and Carlton Streets, Buffalo, NY, USA.
320
Dept of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
321
The Scientific Platforms, the Westmead Institute for Medical Research, the Westmead Research Hub, Westmead, New South Wales, Australia.
322
Flow Cytometry Core Facility, Blizard Institute, Queen Mary London University, London, UK.
323
Faculty of Biological Sciences, Friedrich Schiller University Jena, Jena, Germany.
324
Medinfomatics Ltd., West Drayton, Middlesex, UK.
325
Department for Immunology, Leibniz Research Centre for Working Environment and Human Factors at TU Dortmund (IfADo), Dortmund, Germany.
326
Institute for Cellular & Molecular Immunology, University Medical Center Göttingen, Göttingen, Germany.
327
Department of Infectious Disease, Imperial College London, UK.
328
Wellcome Centre for Infectious Diseases Research in Africa and Department of Medicine, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Republic of South Africa.
329
Tuberculosis Laboratory, The Francis Crick Institute, London, UK.
330
Cooperation Unit for Experimental and Translational Cancer Immunology, Institute of Immunology (Charité - Universitätsmedizin Berlin) and German Cancer Research Center (DKFZ), Berlin, Germany.
331
University Bonn, Medical Faculty, Bonn, Germany.
332
BD Biosciences, San Jose, CA, USA.
333
School of Computing Science, Simon Fraser University, Burnaby, Canada.
334
Department of Rheumatology and Immunology, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
335
Institute of Virology, Technical University of Munich, Munich, Germany.
336
TranslaTUM, Technical University of Munich, Munich, Germany.
337
Maurice Müller Laboratories (Department of Biomedical Research), Universitätsklinik für Viszerale Chirurgie und Medizin Inselspital, University of Bern, Bern, Switzerland.

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.

PMID:
31633216
DOI:
10.1002/eji.201970107

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