{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T04:39:06Z","timestamp":1773290346557,"version":"3.50.1"},"reference-count":18,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2012,8,28]],"date-time":"2012-08-28T00:00:00Z","timestamp":1346112000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2012,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>EdU (5\u2010ethynyl\u20102\u2032\u2010deoxyuridine) incorporation has proved advantageous in the studies of cell kinetics, DNA synthesis, and cellular proliferation <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic> compared to [<jats:sup>3<\/jats:sup>H]thymidine incorporation and BrdU (5\u2010bromo\u20102\u2032\u2010deoxyuridine) incorporation. Here, we describe a method that combines EdU incorporation and immunostaining with flow cytometric analysis to detect the proliferations of T lymphocyte subsets in vitro and optimized the assay's conditions. We found that the number of EdU<jats:sup>+<\/jats:sup> cells were associated with EdU concentration, incubation time, and the volume of Click reaction solution, the best EdU concentration 10\u201350 \u03bcM, the optimal incubation time 8\u201312 h and the proper volume of Click volume 100 \u03bcl for labeling 1 \u00d7 10<jats:sup>6<\/jats:sup> lymphocytes. Fixation was better to be performed before permeabilization, not together with. Furthermore, the permeabilization detergent reagent, PBS with 0.05% saponin was better than Tris buffer saline (TBS) with 0.1% Triton X\u2010100. In addition, sufficient wash with PBS with 0.05% saponin has no influence on the staining of EdU<jats:sup>+<\/jats:sup> cells. Also, the lymphocytes incorporating EdU could be stored at 4\u00b0C, \u221280\u00b0<jats:bold>C,<\/jats:bold> and in liquid nitrogen up to 21 days. The present study will aid in optimization of flow cytometry assay to detect the proliferations of T cell subsets by EdU incorporation and the labeling of cell surface antigens. \u00a9 2012 International Society for Advancement of Cytometry<\/jats:p>","DOI":"10.1002\/cyto.a.22113","type":"journal-article","created":{"date-parts":[[2012,8,28]],"date-time":"2012-08-28T20:03:22Z","timestamp":1346184202000},"page":"901-909","source":"Crossref","is-referenced-by-count":28,"title":["Multicolor flow cytometry analysis of the proliferations of T\u2010lymphocyte subsets <i>in vitro<\/i> by EdU incorporation"],"prefix":"10.1002","volume":"81A","author":[{"given":"Yanli","family":"Sun","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Sun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guigao","family":"Lin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rui","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kuo","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiehong","family":"Xie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lunan","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinming","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2012,8,28]]},"reference":[{"key":"e_1_2_5_2_2","first-page":"718","article-title":"A comparison between bromodeoxyuridine and 3H thymidine labeling in human breast tumors","volume":"4","author":"Waldman FM","year":"1991","journal-title":"Mod Pathol"},{"key":"e_1_2_5_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrn700"},{"key":"e_1_2_5_4_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0712168105"},{"key":"e_1_2_5_5_2","first-page":"e2147","article-title":"Visualization of mitochondrial DNA replication in individual cells by EdU signal amplification","volume":"45","author":"Halnes KM","year":"2010","journal-title":"J Vis Exp"},{"key":"e_1_2_5_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2008.10.006"},{"key":"e_1_2_5_7_2","doi-asserted-by":"publisher","DOI":"10.2144\/000112812"},{"key":"e_1_2_5_8_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20712"},{"key":"e_1_2_5_9_2","doi-asserted-by":"publisher","DOI":"10.1186\/ar2939"},{"key":"e_1_2_5_10_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20914"},{"key":"e_1_2_5_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2009.07.008"},{"key":"e_1_2_5_12_2","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-09-4398"},{"key":"e_1_2_5_13_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI25052"},{"key":"e_1_2_5_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1759(01)00428-8"},{"key":"e_1_2_5_15_2","doi-asserted-by":"publisher","DOI":"10.1007\/s11060-011-0621-6"},{"key":"e_1_2_5_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2008.10.006"},{"key":"e_1_2_5_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2007.07.006"},{"key":"e_1_2_5_18_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20392"},{"key":"e_1_2_5_19_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.990120612"}],"container-title":["Cytometry Part A"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fcyto.a.22113","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.a.22113","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,11]],"date-time":"2023-10-11T02:44:09Z","timestamp":1696992249000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cyto.a.22113"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,8,28]]},"references-count":18,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2012,10]]}},"alternative-id":["10.1002\/cyto.a.22113"],"URL":"https:\/\/doi.org\/10.1002\/cyto.a.22113","archive":["Portico"],"relation":{},"ISSN":["1552-4922","1552-4930"],"issn-type":[{"value":"1552-4922","type":"print"},{"value":"1552-4930","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,8,28]]}}}