{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T06:19:23Z","timestamp":1792131563299,"version":"4.4.0"},"reference-count":72,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2008,9,11]],"date-time":"2008-09-11T00:00:00Z","timestamp":1221091200000},"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":[[2008,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Cell\u2010tracking reagents such as the green\u2010fluorescent protein labeling dye CFSE and the red\u2010fluorescent lipophilic membrane dye PKH26 are commonly used to monitor cell proliferation by flow cytometry in heterogeneous cell populations responding to immune stimuli. Both reagents stain cells with a bright homogeneous fluorescence, which is partitioned between daughter cells during each cell division. Because daughter cell fluorescence intensities are approximately halved after each division, the intensity of a cell relative to its intensity at the time of staining provides information about how many divisions it has undergone. Knowing how many rounds of division have occurred and the relative number of cells in each daughter generation, one can back\u2010calculate the number of cells in the original population (<jats:italic>i.e<\/jats:italic>., cells present at the time of stimulus) that went on to respond by proliferating. Using this information, the precursor cell frequencies and extent of expansion to a specific antigen or mitogen of interest can be calculated. Concurrently, the phenotype of the cells can be determined, as well as their ability to bind antigen or synthesize cytokines, providing more detailed characterization of all cells responding to the antigen, not just effector cells. In multiparameter flow cytometric experiments to simultaneously analyze antigen\u2010specific tetramer binding, cytokine production and T\u2010cell proliferation, we found that only approximately half of the cells that exhibited specific binding to influenza tetramer also proliferated, as measured by dye dilution, and synthesized IFN\u03b3 in response to antigen. We expect the advent of new cell tracking dyes emitting from the violet to the near infrared combined with the increasing number of lasers and detectors on contemporary flow cytometers to further expand the usefulness of this approach to characterization of complex antigen\u2010driven immunological responses. \u00a9 2008 International Society for Advancement of Cytometry<\/jats:p>","DOI":"10.1002\/cyto.a.20619","type":"journal-article","created":{"date-parts":[[2008,9,11]],"date-time":"2008-09-11T16:25:14Z","timestamp":1221150314000},"page":"1019-1034","source":"Crossref","is-referenced-by-count":141,"title":["Tracking antigen\u2010driven responses by flow cytometry: Monitoring proliferation by dye dilution"],"prefix":"10.1002","volume":"73A","author":[{"given":"Paul K.","family":"Wallace","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"suffix":"Jr.","given":"Joseph D.","family":"Tario","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan L.","family":"Fisher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen S.","family":"Wallace","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc S.","family":"Ernstoff","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katharine A.","family":"Muirhead","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2008,10,22]]},"reference":[{"key":"e_1_2_10_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(94)90236-4"},{"key":"e_1_2_10_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2007.297"},{"key":"e_1_2_10_4_2","doi-asserted-by":"publisher","DOI":"10.1080\/08820130701812584"},{"key":"e_1_2_10_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1759(03)00059-0"},{"key":"e_1_2_10_6_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1440-1711.1999.00871.x"},{"key":"e_1_2_10_7_2","doi-asserted-by":"publisher","DOI":"10.1081\/IMM-120038079"},{"key":"e_1_2_10_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(90)90322-M"},{"key":"e_1_2_10_9_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI119873"},{"key":"e_1_2_10_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80605-6"},{"key":"e_1_2_10_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/15256"},{"key":"e_1_2_10_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/79782"},{"key":"e_1_2_10_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/1097-0320(20001015)42:5<277::AID-CYTO4>3.0.CO;2-A"},{"key":"e_1_2_10_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/35065118"},{"key":"e_1_2_10_15_2","doi-asserted-by":"publisher","DOI":"10.1080\/08820130701790368"},{"key":"e_1_2_10_16_2","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-199907270-00024"},{"key":"e_1_2_10_17_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.288.5466.675"},{"key":"e_1_2_10_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/76907"},{"key":"e_1_2_10_19_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2007.296"},{"key":"e_1_2_10_20_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1440-1711.1999.00869.x"},{"key":"e_1_2_10_21_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20097"},{"key":"e_1_2_10_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20437"},{"key":"e_1_2_10_23_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00285-006-0046-6"},{"key":"e_1_2_10_24_2","doi-asserted-by":"publisher","DOI":"10.1186\/1742-4682-4-26"},{"key":"e_1_2_10_25_2","first-page":"S118","article-title":"M.E. 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