Article open access publication

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

European Journal of Immunology, Wiley, ISSN 0014-2980

Volume 49, 10, 2019

DOI:10.1002/eji.201970107, Dimensions: pub.1121951380, PMC: PMC7350392, PMID: 31633216,

Authors

Bari, Sudipto (17) (18)
Barnaba, Vincenzo (19) (20) (21)
Becher, Burkhard (9) (29)
Boulais, Philip E (37) (38)
Brenner, Dirk (40) (41) (42)
Brinkman, Ryan R (33) (43)
Busch, Dirk H (14) (45)
Chow, Sue (57)
Christakou, Eleni (58) (59)
Clerici, Mario (61) (62) (63)
Cook, Laura (33) (34)
Delacher, Michael (72) (73)
Diefenbach, Andreas (2) (68) (77)
Dörner, Thomas (2) (68)
Edinger, Matthias (73) (84)
Feuerer, Markus (72) (73)
Follo, Marie (97) (98)
Frenette, Paul S (37) (38)
Galbraith, David W (103) (104)
Gazzinelli, Ricardo T (106) (107)
Ginhoux, Florent (80) (110) (111)
Goda, Keisuke (112) (113) (114)
Gori, Andrea (119)
Hahn, Jonas (121)
Hansmann, Leo (68) (77) (124)
Hauser, Anja E (2) (77) (127)
Hess, Christoph (64) (130)
Hoffmann, Petra (73) (84)
Höllt, Thomas (91) (132)
Huang, Bo (136)
Hwang, William Y K (17) (18) (140)
Ivison, Sabine M (33) (34)
Jani, Peter K (2) (141)
Keller, Baerbel (145) (146)
Khalilnezhad, Ahad (80) (111)
Kolls, Jay K (148)
Kunkel, Désirée (77) (152)
Kwok, Immanuel (80) (154)
LaRosa, Francesca (61) (62)
Levings, Megan K (33) (34)
Lino, Andreia C (2) (68)
Lopez, Lilly (160)
Luche, Herve (164)
Mantovani, Alberto (169) (170)
Marventano, Ivana (61) (62)
Melchers, Fritz (2) (141)
Mjösberg, Jenny (133) (179)
Moore, Jonni (139)
Moran, Barry (178)
Multhoff, Gabriele (30) (182)
Münz, Christian (9) (29)
Ng, Lai Guan (80) (111) (154) (175) (185)
Ochel, Aaron (184)
Oja, Anna (134)
Piancone, Federica (61) (62)
Piconese, Silvia (20) (21)
Pockley, A Graham (100) (194)
Quinn, Kylie M (36) (196)
Raz, Yotam (201)
Saeys, Yvan (31) (174)
Saresella, Marina (61) (62)
Schadt, Linda (9) (29)
Tan, Leonard (80) (111)
Tárnok, Attila (219) (220) (221)
Tiegs, Gisa (184)
Tornack, Julia (2) (222)
Tree, Timothy I M (58) (59)
Trotter, Joe (217)
Van Gassen, Sofie (31) (174)
Waisman, Ari (231)
Wang, Sa A (234)
Wang, Xin M (235)
Warnatz, Klaus (145) (146)
Warnes, Gary (170)
Warth, Sarah (152)
Waskow, Claudia (198) (236)
Wilkinson, Robert John (69) (240) (241)
Yang, Jennie H M (58) (59)
Yu, Liping (242)
Yue, Alice (243)
Zhao, Yi (244)

Affiliations

Organisations

  1. (1) University of Modena and Reggio Emilia, grid.7548.e
  2. (2) German Rheumatism Research Centre, grid.418217.9
  3. (3) University of Erlangen-Nuremberg, grid.5330.5
  4. (4) Kiel University, grid.9764.c
  5. (5) University Hospital Schleswig-Holstein, grid.412468.d
  6. (6) Lund University, grid.4514.4
  7. (7) Technical University of Denmark, grid.5170.3, DTU
  8. (8) Stanford University, grid.168010.e
  9. (9) University of Zurich, grid.7400.3
  10. (10) Université de Paris, Institut de Recherche Saint-Louis, INSERM U1160, and Gastroenterology Department, Hôpital Saint-Louis - APHP, Paris, France.
  11. (11) University of Lübeck, grid.4562.5
  12. (12) Humanitas Research Hospital, grid.417728.f
  13. (13) University of Birmingham, grid.6572.6
  14. (14) Technical University of Munich, grid.6936.a
  15. (15) University of Florence, grid.8404.8
  16. (16) University of Siena, grid.9024.f
  17. (17) National Cancer Centre Singapore, grid.410724.4
  18. (18) Duke NUS Graduate Medical School, grid.428397.3
  19. (19) Italian Institute of Technology, grid.25786.3e
  20. (20) Istituto Pasteur, grid.452606.3
  21. (21) Sapienza University of Rome, grid.7841.a
  22. (22) Hannover Medical School, grid.10423.34
  23. (23) Fondazione Santa Lucia, grid.417778.a
  24. (24) Medical University of Vienna, grid.22937.3d
  25. (25) University of California, Davis, grid.27860.3b
  26. (26) Ludwig Maximilian University of Munich, grid.5252.0
  27. (27) European Molecular Biology Laboratory, grid.4709.a
  28. (28) University of Technology Sydney, grid.117476.2
  29. (29) Comprehensive Cancer Center Zurich, Switzerland.
  30. (30) Helmholtz Zentrum München, grid.4567.0
  31. (31) Ghent University, grid.5342.0
  32. (32) University College Dublin, grid.7886.1
  33. (33) University of British Columbia, grid.17091.3e
  34. (34) British Columbia Children's Hospital, grid.414137.4
  35. (35) University Hospital Erlangen, grid.411668.c
  36. (36) Monash University, grid.1002.3
  37. (37) Albert Einstein College of Medicine, grid.251993.5
  38. (38) The Ruth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Bronx, New York, USA.
  39. (39) Thermo Fisher Scientific (United States), grid.418190.5
  40. (40) Odense University Hospital, Odense Research Center for Anaphylaxis, University of Southern Denmark, Department of Dermatology and Allergy Center, Odense, Denmark.
  41. (41) University of Luxembourg, grid.16008.3f
  42. (42) Luxembourg Institute of Health, grid.451012.3
  43. (43) BC Cancer Agency, grid.248762.d
  44. (44) University of Auckland, grid.9654.e
  45. (45) German Center for Infection Research, grid.452463.2
  46. (46) Miltenyi Biotec (Germany), grid.59409.31
  47. (47) University of Rochester Medical Center, grid.412750.5
  48. (48) University of Verona, grid.5611.3
  49. (49) University of Melbourne, grid.1008.9
  50. (50) Nankai University, grid.216938.7
  51. (51) University of Gothenburg, grid.8761.8
  52. (52) FIRC Institute of Molecular Oncology, grid.7678.e
  53. (53) Istituto Superiore di Sanità, grid.416651.1
  54. (54) University of Barcelona, grid.5841.8
  55. (55) Paris Descartes University, grid.10992.33
  56. (56) New York University Langone Medical Center, grid.240324.3
  57. (57) Princess Margaret Cancer Centre, grid.415224.4
  58. (58) King's College London, grid.13097.3c
  59. (59) National Institute for Health Research, grid.451056.3
  60. (60) Helmholtz Centre for Infection Research, grid.7490.a
  61. (61) IRCCS Fondazione Don Carlo Gnocchi, Milan, Italy.
  62. (62) University of Milano-Bicocca, grid.7563.7
  63. (63) University of Milan, grid.4708.b
  64. (64) University of Cambridge, grid.5335.0
  65. (65) University of Leicester, grid.9918.9
  66. (66) de Duve Institute, grid.16549.3f
  67. (67) Institut Pasteur, grid.428999.7
  68. (68) Charité, grid.6363.0
  69. (69) The Francis Crick Institute, grid.451388.3
  70. (70) Istituto Giannina Gaslini, grid.419504.d
  71. (71) University of Copenhagen, grid.5254.6, KU
  72. (72) University of Regensburg, grid.7727.5
  73. (73) Regensburg Center for Interventional Immunology (RCI), Regensburg, Germany.
  74. (74) San Raffaele Hospital, grid.18887.3e
  75. (75) Istanbul University, grid.9601.e
  76. (76) Sony (United Kingdom), grid.426400.4
  77. (77) Berlin Institute of Health, grid.484013.a
  78. (78) University of Palermo, grid.10776.37
  79. (79) University of Bonn, grid.10388.32
  80. (80) Singapore Immunology Network, grid.430276.4
  81. (81) Department of Dermatology, Laboratory of Dendritic Cell Biology, Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), University Hospital Erlangen, Erlangen, Germany.
  82. (82) University of Oxford, grid.4991.5
  83. (83) Institute of Immunology, Centre for Infection Medicine, Department of Veterinary Medicine, Freie Universität Berlin, Germany.
  84. (84) University Hospital Regensburg, grid.411941.8
  85. (85) Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Department of Neuropathology, Germany.
  86. (86) University of Toronto, grid.17063.33
  87. (87) Heidelberg Institute for Stem Cell Technology and Experimental Medicine, grid.482664.a
  88. (88) University of Bern, grid.5734.5
  89. (89) Department of Immunology, University L. Eotvos, Budapest, Hungary.
  90. (90) Leibniz Research Institute for Environmental Medicine, Düsseldorf, Germany.
  91. (91) Leiden University Medical Center, grid.10419.3d
  92. (92) Washington University in St. Louis, grid.4367.6
  93. (93) Centro de Investigacion Principe Felipe, grid.418274.c
  94. (94) Newcastle University, grid.1006.7
  95. (95) MDQuest Ltd., Szeged, Hungary.
  96. (96) Necker-Enfants Malades Hospital, grid.412134.1
  97. (97) Department of Medicine I, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
  98. (98) Universitaetsklinikum Freiburg, Lighthouse Core Facility, Zentrum für Translationale Zellforschung, Klinik für Innere Medizin I, Freiburg, Germany.
  99. (99) Owl Biomedical (United States), grid.427089.0
  100. (100) Nottingham Trent University, grid.12361.37
  101. (101) Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Department of Dermatology, Venereology and Allergology.
  102. (102) Leibniz Institute of Photonic Technology, grid.418907.3
  103. (103) Henan University, grid.256922.8
  104. (104) University of Arizona, grid.134563.6
  105. (105) Antoni van Leeuwenhoek Hospital, grid.430814.a
  106. (106) Oswaldo Cruz Foundation, grid.418068.3
  107. (107) University of Massachusetts Medical School, grid.168645.8
  108. (108) Istituto Nazionale Genetica Molecolare, grid.428717.f
  109. (109) University of Veterinary Medicine Vienna, grid.6583.8
  110. (110) Shanghai Jiao Tong University, grid.16821.3c
  111. (111) National University of Singapore, grid.4280.e
  112. (112) University of California, Los Angeles, grid.19006.3e
  113. (113) University of Tokyo, grid.26999.3d
  114. (114) Wuhan University, grid.49470.3e
  115. (115) University of Cologne, grid.6190.e
  116. (116) Alicante Institute for Health and Biomedical Research (ISABIAL), Alicante, Spain.
  117. (117) Networked Biomedical Research Center for Hepatic and Digestive Diseases (CIBERehd), Madrid, Spain.
  118. (118) University of Glasgow, grid.8756.c
  119. (119) Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico, University of Milan.
  120. (120) Roche (United States), grid.418158.1
  121. (121) Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Department of Medicine 3, Rheumatology and Immunology, Universitätsklinikum Erlangen, Erlangen.
  122. (122) Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Zwijnaarde, Belgium.
  123. (123) German Cancer Research Center, grid.7497.d
  124. (124) German Cancer Consortium (DKTK), partner site Berlin, Berlin, Germany.
  125. (125) Department of Medicine and Center for Molecular Medicine at Karolinska University Hospital, Solna, Sweden.
  126. (126) Murdoch Children's Research Institute, grid.1058.c
  127. (127) Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin.
  128. (128) Houston Methodist, grid.63368.38
  129. (129) University of Valencia, grid.5338.d
  130. (130) University Hospital of Basel, grid.410567.1
  131. (131) University of Minnesota, grid.17635.36
  132. (132) Delft University of Technology, grid.5292.c
  133. (133) Karolinska Institute, grid.4714.6
  134. (134) University of Amsterdam, grid.7177.6
  135. (135) New Mexico State University, grid.24805.3b
  136. (136) Chinese Academy of Medical Sciences & Peking Union Medical College, grid.506261.6
  137. (137) Shenzhen University, grid.263488.3
  138. (138) Heidelberg University, grid.7700.0
  139. (139) University of Pennsylvania, grid.25879.31
  140. (140) Singapore General Hospital, grid.163555.1
  141. (141) Max Planck Institute for Infection Biology, grid.418159.0
  142. (142) University of Rijeka, grid.22939.33
  143. (143) Charles University, grid.4491.8
  144. (144) Jena University Hospital, grid.275559.9
  145. (145) Center for Chronic Immunodeficiency, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
  146. (146) Department of Rheumatology and Clinical Immunology, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
  147. (147) ETH Zurich, grid.5801.c
  148. (148) Tulane University, grid.265219.b
  149. (149) Medical University of Graz, grid.11598.34
  150. (150) Goethe University Frankfurt, grid.7839.5
  151. (151) Max Planck Institute for Biology of Ageing, grid.419502.b
  152. (152) BCRT Flow Cytometry Lab, Berlin-Brandenburg Center for Regenerative Therapies, Charité - Universitätsmedizin Berlin.
  153. (153) Osaka University, grid.136593.b
  154. (154) Nanyang Technological University, grid.59025.3b
  155. (155) Institute Curie, grid.418596.7
  156. (156) University of Chieti-Pescara, grid.412451.7
  157. (157) Institut Cochin, CNRS8104, INSERM1016, Department of Endocrinology, Metabolism and Diabetes, Université de Paris, Paris, France.
  158. (158) Caprion Biosciences, ImmuneCarta, Montréal, Quebec, Canada.
  159. (159) Philipp University of Marburg, grid.10253.35
  160. (160) Beckman Coulter (United States), grid.418254.e
  161. (161) IMIM(Hospital de Mar Medical Research Institute), University Pompeu Fabra, Barcelona, Spain.
  162. (162) The Ohio State University, grid.261331.4
  163. (163) Erasmus University Medical Center, grid.5645.2
  164. (164) Aix-Marseille University, grid.5399.6
  165. (165) Canton of St. Gallen-Hospital, grid.413349.8
  166. (166) Heinrich-Pette-Institute, grid.418481.0
  167. (167) Flow and Image Cytometry Shared Resource, Roswell Park Comprehensive Cancer Center, Buffalo, NY, USA.
  168. (168) Fred Hutchinson Cancer Research Center, grid.270240.3
  169. (169) Istituto Clinico Humanitas IRCCS and Humanitas University, Pieve Emanuele, Milan, Italy.
  170. (170) Queen Mary University of London, grid.4868.2
  171. (171) University of Manitoba, grid.21613.37
  172. (172) National Institute of Geriatrics, Rheumatology and Rehabilitation, grid.460480.e
  173. (173) National Research Council, grid.5326.2
  174. (174) Flanders Institute for Biotechnology, grid.11486.3a
  175. (175) University of Sydney, grid.1013.3
  176. (176) Clinical Trial Center Leipzig, University Leipzig, Leipzig, Germany.
  177. (177) Northwestern University, grid.16753.36
  178. (178) Trinity College Dublin, grid.8217.c
  179. (179) Linköping University, grid.5640.7
  180. (180) Bambino Gesù Children's Hospital, grid.414125.7
  181. (181) Helmholtz Centre for Environmental Research, grid.7492.8
  182. (182) Rechts der Isar Hospital, grid.15474.33
  183. (183) Chiba University, grid.136304.3
  184. (184) University Medical Center Hamburg-Eppendorf, grid.13648.38
  185. (185) Institute of Hematology & Blood Diseases Hospital, grid.461843.c
  186. (186) University of Michigan, grid.214458.e
  187. (187) University of Salamanca, grid.11762.33
  188. (188) Burnet Institute, grid.1056.2
  189. (189) Inflammation and Oncology, Research, Amgen Inc, South San Francisco, USA.
  190. (190) University of Greifswald, grid.5603.0
  191. (191) Mahidol University, grid.10223.32
  192. (192) University of Tartu, grid.10939.32
  193. (193) Autonomous University of Barcelona, grid.7080.f
  194. (194) Chromocyte Limited, Electric Works, Sheffield, UK.
  195. (195) University of Udine, grid.5390.f
  196. (196) RMIT University, grid.1017.7
  197. (197) Department of Rheumatology and Clinical Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
  198. (198) Fritz Lipmann Institute, grid.418245.e
  199. (199) Max Delbrück Center for Molecular Medicine, grid.419491.0
  200. (200) Purdue University West Lafayette, grid.169077.e
  201. (201) Groene Hart Ziekenhuis, grid.413370.2
  202. (202) Desatoya LLC, Reno, NV, USA.
  203. (203) Renal Transplant Unit, Division of Internal Medicine, Academic Medical Centre, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.
  204. (204) University of Alberta, grid.17089.37
  205. (205) Cell Biology Unit, IRCCS Ospedale Policlinico San Martino, Genova, Italy.
  206. (206) RIKEN Center for Integrative Medical Sciences, Yokohama, Japan.
  207. (207) University of Lausanne, grid.9851.5
  208. (208) Istituto Neurologico Mediterraneo, grid.419543.e
  209. (209) Centre de Recherches des Cordeliers, Equipe Cancer et Immunité anti-tumorale, Paris, France.
  210. (210) Clinic for Orthopedics and Trauma Surgery, University Hospital Bonn, Bonn, Germany.
  211. (211) Otto-von-Guericke University Magdeburg, grid.5807.a
  212. (212) Translational Research Institute, grid.489335.0
  213. (213) AsedaSciences, West Lafayette, IN, USA.
  214. (214) University of Lisbon, grid.9983.b
  215. (215) University of Brescia, grid.7637.5
  216. (216) BD Life Sciences, Ashland, OR, USA.
  217. (217) BD Life Sciences, San Diego, CA, USA.
  218. (218) National Institutes of Health, grid.94365.3d
  219. (219) Fraunhofer Institute for Cell Therapy and Immunology, grid.418008.5
  220. (220) Leipzig University, grid.9647.c
  221. (221) Tsinghua University, grid.12527.33
  222. (222) BioGenes GmbH, Berlin, Germany.
  223. (223) Novartis (Switzerland), grid.419481.1
  224. (224) Pennsylvania State University, grid.29857.31
  225. (225) Alexander Fleming Biomedical Sciences Research Center, grid.424165.0
  226. (226) University of Sao Paulo, grid.11899.38
  227. (227) Christine Kühne Center for Allergy Research and Education (CK-CARE), Davos, Switzerland.
  228. (228) Department of Infection Biology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nuremberg (FAU), Erlangen, Germany.
  229. (229) BIH Center for Regenerative Therapies (BCRT) Charité Universitätsmedizin Berlin and Berlin Institute of Health, Core Unit ImmunoCheck.
  230. (230) A·P·E-Applied Physics & Electronics, Berlin, Germany.
  231. (231) University Medical Center of the Johannes Gutenberg University Mainz, grid.410607.4
  232. (232) Babraham Institute, grid.418195.0
  233. (233) Roswell Park Comprehensive Cancer Center, Elm and Carlton Streets, Buffalo, NY, USA.
  234. (234) The University of Texas MD Anderson Cancer Center, grid.240145.6
  235. (235) The Scientific Platforms, the Westmead Institute for Medical Research, the Westmead Research Hub, Westmead, New South Wales, Australia.
  236. (236) Friedrich Schiller University Jena, grid.9613.d
  237. (237) Medinfomatics Ltd., West Drayton, Middlesex, UK.
  238. (238) TU Dortmund University, grid.5675.1
  239. (239) University Medical Center Göttingen, grid.411984.1
  240. (240) Imperial College London, grid.7445.2
  241. (241) University of Cape Town, grid.7836.a
  242. (242) BD Biosciences (United States), grid.420052.1
  243. (243) Simon Fraser University, grid.61971.38
  244. (244) Department of Rheumatology and Immunology, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
  245. (245) Maurice Müller Laboratories (Department of Biomedical Research), Universitätsklinik für Viszerale Chirurgie und Medizin Inselspital, University of Bern, Bern, Switzerland.

Description

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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