Abstract
The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers.
| Original language | English |
|---|---|
| Pages (from-to) | 2708-3145 |
| Number of pages | 438 |
| Journal | European Journal of Immunology |
| Volume | 51 |
| Issue number | 12 |
| Early online date | 7 Dec 2021 |
| DOIs | |
| Publication status | Published - Dec 2021 |
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In: European Journal of Immunology, Vol. 51, No. 12, 12.2021, p. 2708-3145.
Research output: Contribution to journal › Article › peer-review
TY - JOUR
T1 - Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)
AU - Cossarizza, Andrea
AU - Chang, Hyun Dong
AU - Radbruch, Andreas
AU - Abrignani, Sergio
AU - Addo, Richard
AU - Akdis, Mübeccel
AU - Andrä, Immanuel
AU - Andreata, Francesco
AU - Annunziato, Francesco
AU - Arranz, Eduardo
AU - Bacher, Petra
AU - Bari, Sudipto
AU - Barnaba, Vincenzo
AU - Barros-Martins, Joana
AU - Baumjohann, Dirk
AU - Beccaria, Cristian G.
AU - Bernardo, David
AU - Boardman, Dominic A.
AU - Borger, Jessica
AU - Böttcher, Chotima
AU - Brockmann, Leonie
AU - Burns, Marie
AU - Busch, Dirk H.
AU - Cameron, Garth
AU - Cammarata, Ilenia
AU - Cassotta, Antonino
AU - Chang, Yinshui
AU - Chirdo, Fernando Gabriel
AU - Christakou, Eleni
AU - Čičin-Šain, Luka
AU - Cook, Laura
AU - Corbett, Alexandra J.
AU - Cornelis, Rebecca
AU - Cosmi, Lorenzo
AU - Davey, Martin S.
AU - De Biasi, Sara
AU - De Simone, Gabriele
AU - del Zotto, Genny
AU - Delacher, Michael
AU - Di Rosa, Francesca
AU - Santo, James Di
AU - Diefenbach, Andreas
AU - Dong, Jun
AU - Dörner, Thomas
AU - Dress, Regine J.
AU - Dutertre, Charles Antoine
AU - Eckle, Sidonia B.G.
AU - Eede, Pascale
AU - Evrard, Maximilien
AU - Falk, Christine S.
AU - Feuerer, Markus
AU - Fillatreau, Simon
AU - Fiz-Lopez, Aida
AU - Follo, Marie
AU - Foulds, Gemma A.
AU - Fröbel, Julia
AU - Gagliani, Nicola
AU - Galletti, Giovanni
AU - Gangaev, Anastasia
AU - Garbi, Natalio
AU - Garrote, José Antonio
AU - Geginat, Jens
AU - Gherardin, Nicholas A.
AU - Gibellini, Lara
AU - Ginhoux, Florent
AU - Godfrey, Dale I.
AU - Gruarin, Paola
AU - Haftmann, Claudia
AU - Hansmann, Leo
AU - Harpur, Christopher M.
AU - Hayday, Adrian C.
AU - Heine, Guido
AU - Hernández, Daniela Carolina
AU - Herrmann, Martin
AU - Hoelsken, Oliver
AU - Huang, Qing
AU - Huber, Samuel
AU - Huber, Johanna E.
AU - Huehn, Jochen
AU - Hundemer, Michael
AU - Hwang, William Y.K.
AU - Iannacone, Matteo
AU - Ivison, Sabine M.
AU - Jäck, Hans Martin
AU - Jani, Peter K.
AU - Keller, Baerbel
AU - Kessler, Nina
AU - Ketelaars, Steven
AU - Knop, Laura
AU - Knopf, Jasmin
AU - Koay, Hui Fern
AU - Kobow, Katja
AU - Kriegsmann, Katharina
AU - Kristyanto, H.
AU - Krueger, Andreas
AU - Kuehne, Jenny F.
AU - Kunze-Schumacher, Heike
AU - Kvistborg, Pia
AU - Kwok, Immanuel
AU - Latorre, Daniela
AU - Lenz, Daniel
AU - Levings, Megan K.
AU - Lino, Andreia C.
AU - Liotta, Francesco
AU - Long, Heather M.
AU - Lugli, Enrico
AU - MacDonald, Katherine N.
AU - Maggi, Laura
AU - Maini, Mala K.
AU - Mair, Florian
AU - Manta, Calin
AU - Manz, Rudolf Armin
AU - Mashreghi, Mir Farzin
AU - Mazzoni, Alessio
AU - McCluskey, James
AU - Mei, Henrik E.
AU - Melchers, Fritz
AU - Melzer, Susanne
AU - Mielenz, Dirk
AU - Monin, Leticia
AU - Moretta, Lorenzo
AU - Multhoff, Gabriele
AU - Muñoz, Luis Enrique
AU - Muñoz-Ruiz, Miguel
AU - Muscate, Franziska
AU - Natalini, Ambra
AU - Neumann, Katrin
AU - Ng, Lai Guan
AU - Niedobitek, Antonia
AU - Niemz, Jana
AU - Almeida, Larissa Nogueira
AU - Notarbartolo, Samuele
AU - Ostendorf, Lennard
AU - Pallett, Laura J.
AU - Patel, Amit A.
AU - Percin, Gulce Itir
AU - Peruzzi, Giovanna
AU - Pinti, Marcello
AU - Pockley, A. Graham
AU - Pracht, Katharina
AU - Prinz, Immo
AU - Pujol-Autonell, Irma
AU - Pulvirenti, Nadia
AU - Quatrini, Linda
AU - Quinn, Kylie M.
AU - Radbruch, Helena
AU - Rhys, Hefin
AU - Rodrigo, Maria B.
AU - Romagnani, Chiara
AU - Saggau, Carina
AU - Sakaguchi, Shimon
AU - Sallusto, Federica
AU - Sanderink, Lieke
AU - Sandrock, Inga
AU - Schauer, Christine
AU - Scheffold, Alexander
AU - Scherer, Hans U.
AU - Schiemann, Matthias
AU - Schildberg, Frank A.
AU - Schober, Kilian
AU - Schoen, Janina
AU - Schuh, Wolfgang
AU - Schüler, Thomas
AU - Schulz, Axel R.
AU - Schulz, Sebastian
AU - Schulze, Julia
AU - Simonetti, Sonia
AU - Singh, Jeeshan
AU - Sitnik, Katarzyna M.
AU - Stark, Regina
AU - Starossom, Sarah
AU - Stehle, Christina
AU - Szelinski, Franziska
AU - Tan, Leonard
AU - Tarnok, Attila
AU - Tornack, Julia
AU - Tree, Timothy I.M.
AU - van Beek, Jasper J.P.
AU - van de Veen, Willem
AU - van Gisbergen, Klaas
AU - Vasco, Chiara
AU - Verheyden, Nikita A.
AU - von Borstel, Anouk
AU - Ward-Hartstonge, Kirsten A.
AU - Warnatz, Klaus
AU - Waskow, Claudia
AU - Wiedemann, Annika
AU - Wilharm, Anneke
AU - Wing, James
AU - Wirz, Oliver
AU - Wittner, Jens
AU - Yang, Jennie H.M.
AU - Yang, Juhao
N1 - Funding Information: Hyun-Dong Chang is supported by the Dr. Rolf M. Schwiete Foundation. Susanne Melzer and Attila Tarnok thank De Novo Software for providing FCS Express. Enrico Lugli is supported by a grant from the Associazione Italiana per la Ricerca sul Cancro (AIRC IG20676). Gabriele De Simone and Giovanni Galletti were supported by Fellowships from the Fondazione Italiana per la Ricerca sul Cancro-Associazione Italiana per la Ricerca sul Cancro (FIRC-AIRC). Jun Dong is supported by Deutsche Forschungsgemeinschft (DFG, German Research Foundation) Projektnummer 389687267 and Chinesisch-Deutsches Zentrum f?r Wissenschaftsf?rderung [Sino-German Center for Research Promotion (SGC)] grant C-0072. Nicola Gagliani, Samuel Huber and Franziska Muscate are supported by DFG fundings: SFB841,GA 2441/3-1, HU 1714/10-1. The tetramer APC-conjugated H-2K (d) HIV-1 gag197-205 AMQMLKETI used in TDS assay for mouse blood T cells was obtained through the NIH Tetramer Facility. Larissa Nogueira Almeida was supported by DFG research grant MA 2273/14-1. Supported by the following grants: AIRC 5X1000 2018 id. 21147 (Lorenzo Moretta); AIRC IG 2017 id. 19920 (Lorenzo Moretta); RC-2020 OPBG (Lorenzo Moretta); AIRC and European Union's Horizon 2020 research and innovation program under the Marie Sk?odowska-Curie grant agreement No 800924 (Linda Quatrini). Dirk Baumjohann was supported by Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Emmy Noether Programme BA 5132/1-2 (252623821) and Germany's Excellence Strategy EXC2151 (390873048). The following formed the writing committees for the indicated sections: A. Graham Pockley, Gemma A. Foulds, Gabriele Multhoff (I.1Essential Controls and Reproducibility in Flow Cytometry); Susanne Melzer, Attila Tarnok (I.2 MIFlowCyt compliance and data display of flow cytometry data in immunological research); Florian Mair, Claudia Haftmann (I.3. Panel design in high-dimensional flow cytometry); Sara De Biasi, Lara Gibellini, Andrea Cossarizza (II. Rare cells ? General rules); Samuele Notarbartolo, Daniela Latorre, Antonino Cassotta, Chiara Vasco, Federica Sallusto, Sergio Abrignani, Jens Geginat (III.1. Human conventional ?? CD4+ T cells); Regina Stark, Kylie Quinn, Jessica Borger (III.2 Murine Conventional ?? CD4+ T cells); Jasper J. P. van Beek, Gabriele De Simone, Giovanni Galletti, Enrico Lugli (III.3 Human conventional ?? CD8 T cells); Regina Stark, Kylie Quinn, Jessica Borger (III.4 Murine Conventional ?? CD8 T cells); Jun Dong (III.5 Human tissue resident memory T cells); Regina Stark, Klaas van Gisbergen (III.6 Murine tissue resident memory T cells); Jos? Antonio Garrote, Aida Fiz, David Bernardo, Fernando Gabriel Chirdo, Eduardo Arranz (III.7 Human intestinal intraepithelial T cells); Immo Prinz, Joana Barros-Martins (III.8 Mouse intestinal intraepithelial T cells); Marcello Pinti (III.9 Immune senescence (aging) of human T cells); Luka ?i?in-?ain, Katarzyna M. Sitnik, Kylie M. Quinn (III.10 Immune senescence (aging) in murine T cells); Dominic A Boardman, Eleni Christakou, Laura Cook, Michael Delacher, Markus Feuerer, Qing Huang, Sabine M Ivison, Megan K Levings, Katherine N. MacDonald, Timothy I.M. Tree, Kirsten A Ward-Hartstonge, Jennie H. M. Yang (III.11 Human FOXP3+ regulatory T cells); Michael Delacher, Juhao Yang, Jana Niemz, Lieke Sanderink, Markus Feuerer, Jochen Huehn (III.12 Murine Foxp3+ regulatory T cells); Chiara Vasco, Nadia Pulvirenti, Paola Gruarin, Jens Geginat (III.13 Human IL-10 producing regulatory T cells (Tr1 cells)); Franziska Muscate, Leonie Brockmann, Samuel Huber, Nicola Gagliani (III.14 Murine Tr1 cells); Anouk von Borstel, Martin S. Davey (III.15 Unconventional human T cells: gamma delta (??) T cells); Immo Prinz, Anneke Wilharm, Inga Sandrock (III.16 Unconventional murine T cells: ?? T cells); Christopher M. Harpur, Dale I. Godfrey, Garth Cameron (III.17 Unconventional human T cells: NKT cells); Andreas Krueger, Heike Kunze-Schumacher, Nikita A. Verheyden (III.18 Unconventional murine T cells: NKT cells); Garth Cameron, Nicholas A. Gherardin, Alexandra J. Corbett, Sidonia B. G. Eckle, James McCluskey, Dale I. Godfrey, Hui-Fern Koay (III.19 Unconventional human T cells: mucosal-Associated Invariant T (MAIT) cells); Andreas Krueger, Heike Kunze-Schumacher, Nikita A. Verheyden (III.20 Unconventional murine T cells: MAIT cells); Carina Saggau, Alexander Scheffold, Petra Bacher (IV.1 Antigen-specific T cell cytometry: Functional read-outs); Anastasia Gangaev, Steven Ketelaars, Pia Kvistborg (IV.2 Measuring antigen-specific CD8 T cell responses); Immanuel Andr?, Dirk Busch, Matthias Schiemann, Kilian Schober (IV.3 Antigen-specific T cell cytometry: MHC multimers); Alessio Mazzoni, Laura Maggi, Francesco Annunziato (IV.4 Identification of SARS-CoV-2 specific human CD4+ T cells by 8-color flow cytometry); Dirk Baumjohann, Yinshui Chang, Johanna E. Huber (IV. 5 Adoptive T cell transfers as a read-out for Ag-specific immune responses in mice); Maria B. Rodrigo, Nina Kessler, Laura Knop, Natalio Garbi, Thomas Sch?ler (IV.6 Cytotoxicity); Alessio Mazzoni, Laura Maggi, Francesco Annunziato (IV.7 Measurement of signal transduction pathways in human T cells); Alessio Mazzoni, Laura Maggi, Lorenzo Cosmi (IV.8 Live cytokine-producing cell sorting with cytokine secretion assayTM); Laura Maggi, Alessio Mazzoni, Francesco Liotta (IV.9 Quantification of soluble cytokines with cytometric bead array); James Wing, Ilenia Cammarata, Vincenzo Barnaba, Shimon Sakaguchi (IV.10 Treg suppression assays); Ambra Natalini, Sonia Simonetti, Giovanna Peruzzi, Miguel Mu?oz-Ruiz, Leticia Monin, Irma Pujol-Autonell, Hefin Rhys, Heather M. Long, Adrian C. Hayday, Francesca Di Rosa (IV.11 Measurement of T cell proliferation by the TDS assay); Annika Wiedemann, Andreia Lino, Thomas D?rner (V.1 Human B cells and their subsets); Annika Wiedemann, Andreia C. Lino, H. Kristyanto, Hans U. Scherer, Thomas D?rner (V.2 Human antigen-specific B cells); Willem van de Veen, Oliver Wirz, M?beccel Akdis (V.3 Human immunoglobulin heavy chain isotypes); Oliver Wirz, Willem van de Veen, M?beccel Akdis (V.4 Human regulatory B cells); Larissa Nogueira Almeida, Rudolf Armin Manz (V.5 Murine B cells and their subsets, including Bregs); Antonia Niedobitek, Axel R. Schulz, Marie Burns, Julia Schulze, Henrik E. Mei (V.6 Human antibody-secreting plasmablasts and plasma cells); Katharina Kriegsmann, Leo Hansmann, Calin Manta, Michael Hundemer (V.7 Human plasma cells in multiple myeloma); Sebastian Schulz, Katharina Pracht, Jens Wittner, Dirk Mielenz, Wolfgang Schuh, Hans-Martin J?ck (V.8 Murine antibody-secreting plasmablasts and plasma cells); Franziska Szelinski, Annika Wiedemann, Andreia C. Lino, Thomas D?rner (VI.1 Measurement of signal transduction pathways in human B cells); Simon Fillatreau, Rudolf Armin Manz, Guido Heine (VI.2 Live cytokine-producing human B cell sorting with secretion assay: the case of IL-10); Baerbel Keller, Marie Follo, Klaus Warnatz (VI.3 Antigen-receptor mediated Ca2+ mobilization in lymphocytes); Dirk Baumjohann, Yinshui Chang, Johanna E. Huber (VI.4 Adoptive B cell transfers as a read-out for Ag-specific immune responses in mice); Daniela Carolina Hern?ndez, Christina Stehle, Oliver Hoelsken, Andreas Diefenbach, James Di Santo, Chiara Romagnani (VII.1 Human Innate Lymphoid Cells); Christina Stehle, Daniela Carolina Hern?ndez, Oliver Hoelsken, Andreas Diefenbach, James Di Santo, Chiara Romagnani (VII.2 Murine Innate Lymphoid Cells); Linda Quatrini, Lorenzo Moretta, Genny del Zotto (VIII.1 Human NK cells); Linda Quatrini, Lorenzo Moretta, Genny del Zotto (VIII.2 Murine NK cells); Regine J. Dress, Amit A. Patel, Charles-Antoine Dutertre, Florent Ginhoux (IX.1 Human mononuclear phagocytes: Monocytes, macrophages, and dendritic cells); Regine J. Dress, Amit A. Patel, Charles-Antoine Dutertre, Florent Ginhoux (IX.2 Murine mononuclear phagocytes: Monocytes, macrophages, and dendritic cells); Regine J. Dress, Amit A. Patel, Charles-Antoine Dutertre, Florent Ginhoux (X.1 Human mononuclear phagocytes: Monocytes, macrophages, and dendritic cells); Regine J. Dress, Amit A. Patel, Charles-Antoine Dutertre, Florent Ginhoux (X.2 Murine mononuclear phagocytes: Monocytes, macrophages, and dendritic cells); Janina Schoen, Christine Schauer, Jeeshan Singh, Jasmin Knopf, Luis Enrique Mu?oz, Martin Herrmann (XI.1 Neutrophil, eosinophil, and basophil phenotypes); Janina Schoen, Christine Schauer, Jeeshan Singh, Jasmin Knopf, Luis Enrique Mu?oz, Martin Herrmann (XI.2 Human bone marrow and cord blood neutrophils); Janina Schoen, Christine Schauer, Jeeshan Singh, Jasmin Knopf, Luis Enrique Mu?oz, Martin Herrmann (XI.3 Murine bone marrow neutrophils); Frank A. Schildberg (XII.1 Human bone marrow stromal cells); Rebecca Cornelis, Richard Addo, Daniel Lenz, Mir-Farzin Mashreghi (XII.2 Murine bone marrow stromal cells); Peter K. Jani, Julia Tornack, Fritz Melchers, Gulce Itir Percin, Julia Fr?bel, Claudia Waskow (XIII Hematopoietic stem cell phenotypes); Jenny F. Kuehne, Christine S. Falk (XIV Tumor cell phenotypes); Lennard Ostendorf, Chotima B?ttcher, Sarah Starossom, Katja Kobow, Pascale Eede, Helena Radbruch (XV Brain and neural cell phenotypes); Laura J. Pallett, Mala K. Maini, Matteo Iannacone (XVI.1 Liver cell phenotypes); Katrin Neumann, Francesco Andreata, Cristian G. Beccaria, Matteo Iannacone (XVI.2 Murine liver cells). Funding Information: Hyun‐Dong Chang is supported by the Dr. Rolf M. Schwiete Foundation. Susanne Melzer and Attila Tarnok thank De Novo Software for providing FCS Express. Enrico Lugli is supported by a grant from the Associazione Italiana per la Ricerca sul Cancro (AIRC IG20676). Gabriele De Simone and Giovanni Galletti were supported by Fellowships from the Fondazione Italiana per la Ricerca sul Cancro‐Associazione Italiana per la Ricerca sul Cancro (FIRC‐AIRC). Jun Dong is supported by Deutsche Forschungsgemeinschft (DFG, German Research Foundation) Projektnummer 389687267 and Chinesisch‐Deutsches Zentrum für Wissenschaftsförderung [Sino‐German Center for Research Promotion (SGC)] grant C‐0072. Nicola Gagliani, Samuel Huber and Franziska Muscate are supported by DFG fundings: SFB841,GA 2441/3‐1, HU 1714/10‐1. The tetramer APC‐conjugated H‐2K (d) HIV‐1 gag AMQMLKETI used in TDS assay for mouse blood T cells was obtained through the NIH Tetramer Facility. Larissa Nogueira Almeida was supported by DFG research grant MA 2273/14‐1. Supported by the following grants: AIRC 5X1000 2018 id. 21147 (Lorenzo Moretta); AIRC IG 2017 id. 19920 (Lorenzo Moretta); RC‐2020 OPBG (Lorenzo Moretta); AIRC and European Union's Horizon 2020 research and innovation program under the Marie Skłodowska‐Curie grant agreement No 800924 (Linda Quatrini). Dirk Baumjohann was supported by Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Emmy Noether Programme BA 5132/1‐2 (252623821) and Germany's Excellence Strategy EXC2151 (390873048). 197‐205 Publisher Copyright: © 2021 Wiley-VCH GmbH
PY - 2021/12
Y1 - 2021/12
N2 - The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers.
AB - The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers.
UR - http://www.scopus.com/inward/record.url?scp=85119620202&partnerID=8YFLogxK
U2 - 10.1002/eji.202170126
DO - 10.1002/eji.202170126
M3 - Article
AN - SCOPUS:85119620202
SN - 0014-2980
VL - 51
SP - 2708
EP - 3145
JO - European Journal of Immunology
JF - European Journal of Immunology
IS - 12
ER -