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

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.

J. Paul Robinson | Jonathan A. Rebhahn | Kylie M. Quinn | F. Ginhoux | V. Beneš | Y. Saeys | B. Becher | N. Aghaeepour | D. Busch | H. Hammad | S. Müller | A. Schlitzer | P. Klenerman | D. Voehringer | A. Cumano | Bartek Rajwa | D. Galbraith | J. Trowsdale | A. Scheffold | S. Kaufmann | H. Rahn | T. Kalina | A. Órfão | K. Goda | A. Cooke | M. Veldhoen | A. Oxenius | B. Engelhardt | H. Ljunggren | A. Mantovani | S. Jonjić | R. Brinkman | M. van den Broek | W. Ouyang | P. Wallace | C. Sautès-Fridman | A. Waisman | P. Chattopadhyay | M. Clerici | Michael Loran Dustin | G. Matarese | Hyun-Dong Chang | D. Hedley | J. O'connor | H. Jäck | D. Speiser | K. Takeda | H. Ulrich | T. Höfer | M. Feuerer | N. Stanley | B. Gaudillière | V. Barnaba | S. Piconese | D. Scott‐Algara | T. Mosmann | H. Maecker | S. Van Gassen | B. Ludewig | A. Filby | C. Dutertre | Immanuel Kwok | L. Ng | T. Höllt | A. Iannone | T. Winkler | Bianka Baying | J. McCluskey | S. Hartmann | E. Remmerswaal | R. V. van Lier | K. Hogquist | H. Volk | M. Mcgrath | R. Holmdahl | G. Willimsky | D. Schraivogel | P. Coulie | J. Houston | B. Everts | W. Pickl | C. Hess | H. Stockinger | R. Wilkinson | P. Peterson | T. Korn | J. González-Navajas | P. Frenette | J. Ruland | L. Cosmi | F. Annunziato | M. Akdiş | T. Dörner | S. Nourshargh | A. Ståhlberg | S. Miller | L. Chatenoud | T. Kurosaki | O. Lantz | J. D. Di Santo | G. Borsellino | L. Battistini | C. Bogdan | C. Haftmann | D. Brenner | M. Lohoff | C. Esser | Liping Yu | I. Panse | A. Radbruch | Y. Samstag | Y. Takahama | A. Krueger | A. Celada | G. Anderson | D. Sester | R. Gazzinelli | A. Santoni | M. López-Botet | J. Kolls | A. Zychlinsky | W. Maśliński | I. McInnes | P. Wurst | D. Davies | Xuetao Cao | G. Van Isterdael | A. Tárnok | C. Baldari | S. Leibundgut-Landmann | M. Levings | S. Sozzani | G. Hansson | F. Schildberg | Guadalupe Herrera | A. Martínez-Romero | K. Warnatz | F. Melchers | C. Pucillo | F. Ebner | M. Yazdanbakhsh | Takashi Saito | V. D. Dang | S. Fillatreau | F. Dieli | T. Nakayama | G. Tiegs | M. Kopf | R. Beyaert | A. Stall | S. Casola | K. Mills | B. Hoyer | S. Ziegler | M. Paulsen | A. Hauser | J. Petriz | A. Anselmo | Z. Poon | L. Čičin-Šain | Michael Delacher | C. Kurts | G. Hämmerling | R. Palankar | N. Baumgarth | P. Bacher | L. Moretta | S. Sakaguchi | D. Mielenz | H. Koay | D. Godfrey | G. Multhoff | M. Saresella | I. Marventano | F. Piancone | F. Mair | K. Kobow | D. Pavlinić | R. Dress | Ahad Khalilnezhad | A. Cossarizza | A. Acs | D. Adam | S. Adam‐Klages | W. Agace | M. Allez | L. N. Almeida | Giorgia Alvisi | I. Andrä | S. Bari | J. Barros-Martins | W. Bauer | S. Baumgart | Dirk Baumjohann | M. Bebawy | W. Beisker | A. Blanco | D. Boardman | J. Borger | P. Boulais | Jolene A. Bradford | A. Brooks | M. Büscher | T. Bushnell | F. Calzetti | G. Cameron | Ilenia Cammarata | S. Cardell | M. Cassatella | A. Cavani | S. Chow | Eleni Christakou | F. Colombo | L. Cook | A. Cooper | A. Corbett | A. Cosma | L. Cvetkovic | C. Dang-Heine | M. Davey | S. De Biasi | Genny Del Zotto | G. D. Dela Cruz | S. Della Bella | P. Dellabona | G. Deniz | M. Dessing | A. Diefenbach | A. Dolf | D. Dudziak | S. Eckle | M. Edinger | P. Eede | G. R. Ehrhardt | Marcus Eich | P. Engel | A. Erdei | M. Evrard | C. Falk | T. Fehniger | Mar Felipo-Benavent | H. Ferry | Kata Filkor | M. Follo | I. Förster | John Foster | G. Foulds | Britta Frehse | S. Frischbutter | W. Fritzsche | A. Gangaev | N. Garbi | J. Geginat | W. Gerner | N. Gherardin | K. Ghoreschi | L. Gibellini | Christoph Goettlinger | C. Goodyear | A. Gori | J. Grogan | D. Grummitt | A. Grützkau | J. Hahn | L. Hansmann | C. Harpur | A. Hauser | D. Haviland | Daniela C Hernández | M. Herrmann | P. Hoffmann | Tristan Holland | P. Hombrink | Bo Huang | F. Huang | Johanna E. Huber | J. Huehn | M. Hundemer | Christopher A Hunter | W. Hwang | Florian Ingelfinger | S. Ivison | P. Jani | B. Jávega | Toralf Kaiser | T. Kamradt | Baerbel Keller | S. Ketelaars | Srijit Khan | J. Kisielow | J. Knopf | Wan Ting Kong | K. Kriegsmann | H. Kristyanto | Thomas Kroneis | J. Kühne | C. Kukat | D. Kunkel | H. Kunze-Schumacher | P. Kvistborg | Jonathan J. M. Landry | P. Lanuti | Francesca Larosa | A. Lehuen | M. Leipold | Leslie Y T Leung | A. Lino | F. Liotta | V. Litwin | Yanling Liu | G. Lombardi | L. Lopez | A. Lovett-racke | E. Lubberts | H. Luche | E. Lugli | S. Lunemann | L. Maggi | O. Maguire | K. Mair | R. Manz | A. Marshall | G. Martrus | A. Mattioli | C. Maueröder | A. Mazzoni | H. McGuire | H. Mei | S. Melzer | H. Minderman | J. Mjösberg | Jonni S. Moore | B. Moran | L. Munoz | C. Münz | M. Nasi | K. Neumann | Antonia Niedobitek | Gabriel Núñez | A. Ochel | A. Oja | Diana Ordonez | Eva Orlowski-Oliver | K. Pattanapanyasat | L. Penter | C. Peth | M. Pinti | A. Pockley | M. Podolska | Katharina Pracht | I. Prinz | S. Quataert | L. Quatrini | H. Radbruch | T. Radstake | S. Rahmig | G. Ravichandran | Y. Raz | D. Recktenwald | D. Reimer | C. Reis e Sousa | L. Richter | Laura G. Rico | A. Riddell | A. Rieger | C. Romagnani | A. Rubartelli | A. Saalmüller | F. Sala-de-Oyanguren | S. Sanderson | I. Sandrock | R. Sanz | B. Sawitzki | Linda Schadt | H. Scherer | M. Schiemann | Esther Schimisky | J. Schlosser | Stephan A Schmid | S. Schmitt | K. Schober | W. Schuh | T. Schüler | R. Schulte | A. Schulz | Sebastian R. Schulz | C. Scottà | T. Shankey | B. Silva-Santos | A. Simon | K. Sitnik | Josef Spidlen | R. Stark | Christina Stehle | Tobit Steinmetz | Leonard Tan | G. Toldi | J. Tornack | E. Traggiai | M. Trebak | T. Tree | J. Trotter | M. Tsoumakidou | Sophia Urbanczyk | W. van de Veen | E. van der Pol | Salvador Vento-Asturias | P. Vieira | A. von Borstel | K. von Volkmann | Rachael V Walker | Sa A. Wang | X. Wang | Mike D. Ward | Kirsten A. Ward-Hartstonge | G. Warnes | Sarah Warth | C. Waskow | J. Watson | C. Watzl | Leonie Wegener | T. Weisenburger | A. Wiedemann | J. Wienands | A. Wilharm | J. Wing | Rieke Winkelmann | O. Wirz | Alicia Y. W. Wong | Jennie H M Yang | Juhao Yang | Alice Yue | Hanlin Zhang | Yi Zhao | C. Zielinski | J. Zimmermann | L. Cvetković | W. Veen | Götz R. A. Ehrhardt | Jennie H. M. Yang | R. V. Lier | G. Anderson | V. Dang | Pablo Engel | J. Landry | Hanlin Zhang | A. Mattioli | M. Broek | Christopher M. Harpur | Leslie Y. T. Leung | Michael L. Dustin | Tobit D. Steinmetz | Christine Falk | Christopher A. Hunter | Francisco Sala-de-Oyanguren | F. Ingelfinger | Kilian Schober | T. Kroneis | Jonni Moore | M. McGrath | Lisa Richter | Jasmin Knopf | W. Maslinski | G. Ehrhardt | Dorothea Reimer | Pleun Hombrink | T. Saito | Paola Lanuti | Andrea Gori | Stephan A. Schmid | Fritz Melchers | Joana Barros-Martins | T. Vincent Shankey | Anne Cooke | J. Wienands | Marcello Pinti | Andrea Cossarizza | Günter Hämmerling | Todd A. Fehniger | Marco A. Cassatella | Frank A. Schildberg | Ryan R. Brinkman | Winfried F. Pickl | Annavittoria Mattioli | Thomas Weisenburger | Daniel Scott‐Algara | Arturo Zychlinsky | Gemma A. Foulds | Andreas Schlitzer | Cosima T. Baldari | Florent Ginhoux | Diether Recktenwald | Susanna L. Cardell | Chiara Romagnani | Silvano Sozzani | Virginia Litwin | Jonas Hahn | Anna Rubartelli | Liping Yu | Henning Ulrich | Thomas Höfer | Milena Nasi | Anna Vittoria Mattioli | Frank A. Schildberg | Antonio Celada | Enrico Lugli | Jenny Mjösberg | Amy E. Lovett‐Racke | B. Gaudilliere | Frank A Schildberg | D. Ordonez | Livius Penter | Jakob Zimmermann | Irmgard Förster | Antonio Cosma | Paul S. Frenette | Caetano Reis e Sousa | Sally A. Quataert | Regina Stark | Lisa Richter | William W. Agace | Dominic A. Boardman | Jürgen Ruland | Winfried F. Pickl | Hannes Stockinger | T. Radstake | Manfred Kopf | W. Maśliński | Gabriele Multhoff | Carl S. Goodyear | Andrea Cossarizza | Francesco Annunziato | Lorenzo Cosmi | Francesco Liotta | Rikard Holmdahl | Kilian Schober | Christian Bogdan | Dale I. Godfrey | T. Saito | Giovanna Lombardi | Alberto Orfao | Giovanna Lombardi | Maria Yazdanbakhsh | Andrea Gori | Yi Zhao | Birgit Sawitzki | J. Paul Robinson | James Di Santo | S. van Gassen | Armin Saalmüller | Christian Maueröder | Kiyoshi Takeda | M. Yazdanbakhsh | Bart Everts | Helena Radbruch | Pratip K. Chattopadhyay | Miguel López-Botet | Thomas Höfer | Arturo Zychlinsky | H. T. Maecker | Xuetao Cao | Alberto Mantovani | Silvano Sozzani | Angela Santoni | Andreas Diefenbach | Dirk Brenner | John Trowsdale | Yousuke Takahama | Toshinori Nakayama | William W. Agace | J. K. Kolls | H M McGuire | Ana Cumano | Anna E. Oja | Susann Rahmig | Ari Waisman | Günter Hämmerling | A. G. Pockley | Keisuke Goda | G. Warnes | Mario Clerici | Pärt Peterson | Dirk Busch | Jane L. Grogan | Anna Erdei | Hyun-Dong Chang | Andreas Radbruch | Andreas Acs | Dieter Adam | Sabine Adam‐Klages | Mübeccel Akdis | Matthieu Allez | Larissa Nogueira Almeida | Giorgia Alvisi | Graham Anderson | Immanuel Andrä | Achille Anselmo | Petra Bacher | Sudipto Bari | Vincenzo Barnaba | Joana Barros‐Martins | Wolfgang Bauer | Sabine Baumgart | Nicole Baumgarth | Dirk Baumjohann | Bianka Baying | Mary Bebawy | Burkhard Becher | Wolfgang Beisker | Vladimir Benes | Alfonso Blanco | Dominic A. Boardman | Christian Bogdan | Jessica G. Borger | Philip E. Boulais | Jolene A. Bradford | Dirk Brenner | Anna E S Brooks | Martin Büscher | Timothy P. Bushnell | Federica Calzetti | Garth Cameron | Ilenia Cammarata | Stefano Casola | Andrea Cavani | Sue Chow | Eleni Christakou | Luka Čičin‐Šain | Mario Clerici | Laura Cook | Andrea M. Cooper | Alexandra J. Corbett | Antonio Cosma | Pierre G. Coulie | Ana Cumano | Ljiljana Cvetkovic | Chantip Dang‐Heine | Martin S. Davey | Derek Davies | Sara De Biasi | Genny Del Zotto | Gelo Victoriano Dela Cruz | Michael Delacher | Silvia Della Bella | Paolo Dellabona | Günnur Deniz | Mark Dessing | James P. Di Santo | Andreas Diefenbach | Andreas Dolf | T. Dörner | Regine J. Dress | Michael Dustin | Charles‐Antoine Dutertre | Friederike Ebner | Sidonia B. G. Eckle | Matthias Edinger | Pascale Eede | Götz R.A. Ehrhardt | Marcus Eich | Pablo Engel | Britta Engelhardt | Anna Erdei | Charlotte Esser | Bart Everts | Maximilien Evrard | Christine S. Falk | Mar Felipo‐Benavent | Helen Ferry | Markus Feuerer | Andrew Filby | Kata Filkor | Simon Fillatreau | Marie Follo | John Foster | Britta Frehse | Stefan Frischbutter | Wolfgang Fritzsche | David W. Galbraith | Anastasia Gangaev | Natalio Garbi | Brice Gaudilliere | Ricardo T. Gazzinelli | Jens Geginat | Wilhelm Gerner | Nicholas A. Gherardin | K. Ghoreschi | Lara Gibellini | Dale I. Godfrey | Christoph Goettlinger | Jose M. González‐Navajas | Daryl Grummitt | Andreas Grützkau | Claudia Haftmann | Hamida Hammad | Göran K. Hansson | Christopher M. Harpur | Susanne Hartmann | Andrea Hauser | Anja E Hauser | David W. Hedley | Martin Herrmann | Christoph Hess | Kristin Hogquist | Tristan Holland | Thomas Höllt | Bimba F. Hoyer | Fang‐Ping Huang | Jochen Huehn | Michael Hundemer | Christopher A. Hunter | William Y. K. Hwang | Anna Iannone | Florian Ingelfinger | Sabine M Ivison | Hans‐Martin Jäck | Peter K. Jani | Beatriz Jávega | Stipan Jonjic | Tomas Kalina | Stefan H. E. Kaufmann | Steven L. C. Ketelaars | Ahad Khalilnezhad | Srijit Khan | Jan Kisielow | P. Klenerman | Hui‐Fern Koay | Katja Kobow | Thomas Korn | Katharina Kriegsmann | Hendy Kristyanto | Thomas Kroneis | Andreas Krueger | Jenny Kühne | Christian Kukat | Désirée Kunkel | Heike Kunze‐Schumacher | Tomohiro Kurosaki | Christian Kurts | Immanuel Kwok | Jonathan Landry | Olivier Lantz | Francesca LaRosa | Agnès Lehuen | Salomé LeibundGut‐Landmann | Michael D. Leipold | Megan K. Levings | Andreia C. Lino | Hans‐Gustaf Ljunggren | Lilly Lopez | Miguel López‐Botet | Hérve Luche | Burkhard Ludewig | Sebastian Lunemann | Holden T. Maecker | Laura Maggi | Orla Maguire | Florian Mair | Kerstin H. Mair | Alberto Mantovani | Rudolf A. Manz | Aaron Marshall | Alicia Martínez‐Romero | Glòria Martrus | Ivana Marventano | Giuseppe Matarese | Christian Maueröder | James McCluskey | Helen M. McGuire | Iain B. McInnes | Henrik E. Mei | Susanne Melzer | Dirk Mielenz | Stephen D. Miller | Tim R. Mosmann | Susann Müller | Gabriele Multhoff | Luis Enrique Muñoz | Christian Münz | Toshinori Nakayama | Antonia Niedobitek | José‐Enrique O'Connor | Aaron Ochel | Anna Oja | Diana Ordonez | Eva Orlowski‐Oliver | Annette Oxenius | Raghavendra Palankar | Isabel Panse | Dinko Pavlinic | Livius Penter | Christian Peth | Jordi Petriz | Federica Piancone | A. Graham Pockley | Malgorzata Justyna Podolska | Zhiyong Poon | Katharina Pracht | Immo Prinz | Carlo E. M. Pucillo | Linda Quatrini | Kylie M. Quinn | Tim R. D. J. Radstake | Susann Rahmig | Hans‐Peter Rahn | Bartek Rajwa | Yotam Raz | Jonathan A. Rebhahn | Laura G. Rico | Andy Riddell | Armin Saalmüller | Yvan Saeys | T. Saito | Francisco Sala‐de‐Oyanguren | Sharon Sanderson | Inga Sandrock | Angela Santoni | Ramon Bellmàs Sanz | Marina Saresella | Catherine Sautes‐Fridman | Linda Schadt | Alexander Scheffold | Matthias Schiemann | Esther Schimisky | Josephine Schlosser | Stephan Schmid | Steffen Schmitt | Wolfgang Schuh | Thomas Schüler | Reiner Schulte | Cristiano Scottá | Daniel Scott‐Algara | David P. Sester | Bruno Silva‐Santos | Anna Katharina Simon | Katarzyna M. Sitnik | Daniel E. Speiser | Josef Spidlen | Anders Stahlberg | Natalie Stanley | Tobit Steinmetz | Yousuke Takahama | Leonard Tan | Attila Tárnok | Gisa Tiegs | Gergely Toldi | Julia Tornack | Elisabetta Traggiai | Mohamed Trebak | Timothy I.M. Tree | Joe Trotter | Maria Tsoumakidou | Willem Veen | Maries Broek | Edwin Pol | Sofie Van Gassen | Gert Van Isterdael | Marc Veldhoen | Salvador Vento‐Asturias | Paulo Vieira | David Voehringer | Hans-Dieter Volk | Anouk Borstel | Konrad Volkmann | Ari Waisman | Rachael V. Walker | Paul K. Wallace | Sa A. Wang | Xin M. Wang | Kirsten A Ward‐Hartstonge | Klaus Warnatz | Gary Warnes | Sarah Warth | Claudia Waskow | James V. Watson | Carsten Watzl | Leonie Wegener | Annika Wiedemann | Anneke Wilharm | Robert J. Wilkinson | Gerald Willimsky | Rieke Winkelmann | Thomas H. Winkler | Oliver F. Wirz | Peter Wurst | Juhao Yang | Alice Yue | Susanne Maria Ziegler | Christina Zielinski | Maximilien Evrard | Kristin A. Hogquist | Francesca LaRosa | Immanuel Andrä | Michael D. Leipold | Francisco Sala‐de‐Oyanguren | Salvador Vento‐Asturias | Konrad v. Volkmann | Claudia Waskow | Sabine Adam‐Klages | Larissa Nogueira Almeida | Petra Bacher | W. Beisker | Vladimir Benes | Jessica G. Borger | Timothy Bushnell | Xuetao Cao | Susanna L. Cardell | Antonio Celada | Pratip K. Chattopadhyay | Eleni Christakou | Anne Cooke | Antonio Cosma | Pierre G. Coulie | C. Dang‐Heine | Britta Engelhardt | Helen Ferry | Irmgard Förster | John Foster | Hamida Hammad | Susanne Hartmann | Andrea Hauser | Bimba Hoyer | Tomáš Kalina | Katja Kobow | Thomas Kroneis | Desiree Kunkel | Heike Kunze-Schumacher | Olivier Lantz | Fritz Melchers | Jonni Moore | Christian Münz | Diana Ordonez | Eva Orlowski‐Oliver | Isabel Panse | Christian Peth | Kylie M. Quinn | Andy Riddell | J. P. Robinson | Anna Rubartelli | Jurgen Ruland | Angela Santoni | Ramon Bellmàs Sanz | Josephine Schlosser | T. Shankey | Anders Stahlberg | A. Tárnok | Mohamed Trebak | Henning Ulrich | Edwin Pol | Paulo Vieira | A. Borstel | Sa A Wang | Xin M. Wang | Klaus Warnatz | Carsten Watzl | Thomas Weisenburger | Rieke Winkelmann | Liping Yu | Hanlin Zhang | Iain B McInnes | Christoph Hess | Sa A. Wang | D. I. Godfrey | Bimba F. Hoyer | Anja E. Hauser | David W Galbraith | Daniel Schraivogel | Tomas Kalina | Vladimír Beneš | Susanna Cardell | Jessica G Borger | Pierre Coulie | J. Kolls | M. Kopf | F. Schildberg | Beatriz Jávega | José-Enrique O’Connor | Anders Ståhlberg

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