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Markers present in the OMIP allow the profiling of neutrophil subsets associated with ontogeny, migration, phagocytosis capacity, granule release, and immune modulation. For panel design, we ensured that the commonly available fluorophores FITC\/AF488, PE, and APC were assigned to the intracellular subset marker Olfactomedin 4, the maturity and activation marker CD10, and whole blood subset marker CD177, respectively. These markers can be easily replaced without affecting the core identification of neutrophils, enabling antibodies to new neutrophil antigens of interest or for fluorescent substrates to assess different neutrophil functions to be easily explored. Panel optimization was performed on whole blood and purified neutrophils. We demonstrate applications on clinical samples (whole blood and saliva) and experimental endpoints (purified neutrophils stimulated through an in vitro transmigration assay). We hope that providing a uniform platform to analyze neutrophil plasticity in various sample types will facilitate the future understanding of neutrophil subsets in health and disease.<\/jats:p>","DOI":"10.1002\/cyto.a.24820","type":"journal-article","created":{"date-parts":[[2024,1,5]],"date-time":"2024-01-05T04:44:59Z","timestamp":1704429899000},"page":"81-87","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["OMIP\u2010100: A flow cytometry panel to investigate human neutrophil subsets"],"prefix":"10.1002","volume":"105","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9771-3194","authenticated-orcid":false,"given":"Craig J.","family":"Schofield","sequence":"first","affiliation":[{"name":"Wal\u2010Yan Respiratory Research Centre, Telethon Kids Institute University of Western Australia  Nedlands Western Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3526-9985","authenticated-orcid":false,"given":"Rabindra","family":"Tirouvanziam","sequence":"additional","affiliation":[{"name":"Department of Pediatrics Emory University  Atlanta Georgia USA"},{"name":"Center for CF &amp; Airways Disease Research Children's Healthcare of Atlanta  Atlanta Georgia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9381-326X","authenticated-orcid":false,"given":"Luke W.","family":"Garratt","sequence":"additional","affiliation":[{"name":"Wal\u2010Yan Respiratory Research Centre, Telethon Kids Institute University of Western Australia  Nedlands Western Australia Australia"},{"name":"Medical School University of Western Australia  Crawley Western Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,1,5]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/eci.12967"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1092385"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21010287"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1500312"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI57990"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1165\/rcmb.2017-0021OC"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells10092406"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0712386105"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1164\/rccm.201803-0442OC"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.08.001"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.4A0915-442RR"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1177\/0022034516645564"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1164\/rccm.201606-1143OC"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2023.105948"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1097\/SHK.0b013e3181e7e61b"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.3001201"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1100450"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciimmunol.aaf8943"},{"issue":"1","key":"e_1_2_8_20_1","article-title":"The immunosuppressive phenotype of tumor\u2010infiltrating neutrophils is associated with obesity in kidney cancer patients","volume":"9","author":"Margaroli C","year":"2020","journal-title":"Onco Targets Ther"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-08-1921"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature15367"},{"issue":"9","key":"e_1_2_8_23_1","first-page":"657","article-title":"Flow cytometric analysis of human bone marrow. 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