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

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.

SUBMITTER: Cossarizza A

PROVIDER: S-EPMC7350392 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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

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

European journal of immunology 20191001 10

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, feat ...[more]

PMID: 31633216

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Project description:Electronic cell sorting for isolation and culture of dinoflagellates and other marine eukaryotic phytoplankton was compared to the traditional method of manually picking cells using a micropipette. Trauma to electronically sorted cells was not a limiting factor, as fragile dinoflagellates, such as Karenia brevis (Dinophyceae), survived electronic cell sorting to yield viable cells. The rate of successful isolation of large-scale (> 4 litres) cultures was higher for manual picking than for electronic cell sorting (2% vs 0.5%, respectively). However, manual picking of cells is more labor intensive and time consuming. Most manually isolated cells required repicking, as the cultures were determined not to be unialgal after a single round of isolation; whereas, no cultures obtained in this study from electronic single-cell sorting required resorting. A broad flow cytometric gating logic was employed to enhance species diversity. The percentages of unique genotypes produced by manual picking or electronic cell sorting were similar (57% vs 54%, respectively), and each approach produced a variety of dinoflagellate or raphidophyte genera. Alternatively, a highly restrictive gating logic was successfully used to target K. brevis from a natural bloom sample. Direct electronic single-cell sorting was more successful than utilizing a pre-enrichment sort followed by electronic single-cell sorting. The appropriate recovery medium may enhance the rate of successful isolations. Seventy percent of isolated cells were recovered in a new medium (RE) reported here, which was optimized for axenic dinoflagellate cultures. The greatest limiting factor to the throughput of electronic cell sorting is the need for manual postsort culture maintenance and assessment of the large number of isolated cells. However, when combined with newly developed automated methods for growth screening, electronic single-cell sorting has the potential to accelerate the discovery of new algal strains.

Dataset Information

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

Publications

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

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