{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T05:01:52Z","timestamp":1788930112872,"version":"build-2803163510"},"reference-count":44,"publisher":"Oxford University Press (OUP)","issue":"12","license":[{"start":{"date-parts":[[2025,2,26]],"date-time":"2025-02-26T00:00:00Z","timestamp":1740528000000},"content-version":"vor","delay-in-days":1366,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["WT109965MA"],"award-info":[{"award-number":["WT109965MA"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]},{"name":"NIHR Senior Fellowship"},{"name":"John Fell Fund"},{"DOI":"10.13039\/501100003249","name":"Laidlaw Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003249","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Human Immune Discovery Initiative Oxford"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,6,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>ChipCytometry is a multiplex imaging method that can be used to analyze either cell suspensions or tissue sections. Images are acquired by iterative cycles of immunostaining with fluorescently labeled Abs, followed by photobleaching, which allows the accumulation of multiple markers on a single sample. In this study, we explored the feasibility of using ChipCytometry to identify and phenotype cell subsets, including rare cell types, using a combination of tissue sections and single-cell suspensions. Using ChipCytometry of tissue sections, we successfully demonstrated the architecture of human palatine tonsils, including the B and T cell zones, and characterized subcompartments such as the B cell mantle and germinal center zone, as well as intrafollicular PD1-expressing CD4+ T cells. Additionally, we were able to identify the rare tonsillar T cell subsets, mucosal-associated invariant T (MAIT) and \u03b3\u03b4-T cells, within tonsil tissue. Using single-cell suspension ChipCytometry, we further dissected human tonsillar T cell subsets via unsupervised clustering analysis as well as supervised traditional manual gating. We were able to show that PD1+CD4+ T cells are comprised of CXCR5+BCL6high follicular Th cells and CXCR5\u2212BCL6mid pre\u2013follicular Th cells. Both supervised and unsupervised analysis approaches identified MAIT cells in single-cell suspensions, confirming a phenotype similar to that of blood-derived MAIT cells. In this study, we demonstrate that ChipCytometry is a viable method for single-cell suspension cytometry and analysis, with the additional benefit of allowing phenotyping in a spatial context using tissue sections.<\/jats:p>","DOI":"10.4049\/jimmunol.2100063","type":"journal-article","created":{"date-parts":[[2021,6,7]],"date-time":"2021-06-07T18:45:44Z","timestamp":1623091544000},"page":"3073-3082","source":"Crossref","is-referenced-by-count":14,"title":["Defining T Cell Subsets in Human Tonsils Using ChipCytometry"],"prefix":"10.1093","volume":"206","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9952-1812","authenticated-orcid":false,"given":"Joachim P","family":"Hagel","sequence":"first","affiliation":[{"name":"Peter Medawar Building for Pathogen Research, Nuffield Department of Medicine, University of Oxford , Oxford ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0220-9097","authenticated-orcid":false,"given":"Kyle","family":"Bennett","sequence":"additional","affiliation":[{"name":"Peter Medawar Building for Pathogen Research, Nuffield Department of Medicine, University of Oxford , Oxford ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0409-406X","authenticated-orcid":false,"given":"Francesca","family":"Buffa","sequence":"additional","affiliation":[{"name":"Computational Biology and Integrative Genomics, Department of Oncology, University of Oxford , Oxford ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4307-9161","authenticated-orcid":false,"given":"Paul","family":"Klenerman","sequence":"additional","affiliation":[{"name":"Peter Medawar Building for Pathogen Research, Nuffield Department of Medicine, University of Oxford , Oxford ,","place":["United Kingdom"]},{"name":"NIHR Biomedical Research Centre, John Radcliffe Hospital, Oxford University Hospitals NHS Foundation Trust , Oxford ,","place":["United Kingdom"]},{"name":"Translational Gastroenterology Unit, Nuffield Department of Medicine, University of Oxford , Oxford ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5299-9344","authenticated-orcid":false,"given":"Christian B","family":"Willberg","sequence":"additional","affiliation":[{"name":"Peter Medawar Building for Pathogen Research, Nuffield Department of Medicine, University of Oxford , Oxford ,","place":["United Kingdom"]},{"name":"NIHR Biomedical Research Centre, John Radcliffe Hospital, Oxford University Hospitals NHS Foundation Trust , Oxford ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1826-953X","authenticated-orcid":false,"given":"Kate","family":"Powell","sequence":"additional","affiliation":[{"name":"Peter Medawar Building for Pathogen Research, Nuffield Department of Medicine, University of Oxford , Oxford ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2021,6,15]]},"reference":[{"key":"2025030417503977700_r1","doi-asserted-by":"crossref","first-page":"43","DOI":"10.29245\/2578-2967\/2018\/1.1122","article-title":"Novel platforms of multiplexed immunofluorescence for study of paraffin tumor tissues","volume":"2","author":"Parra","year":"2018","journal-title":"J. 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