{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,24]],"date-time":"2025-10-24T12:51:29Z","timestamp":1761310289286},"reference-count":19,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2005,9,14]],"date-time":"2005-09-14T00:00:00Z","timestamp":1126656000000},"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":[[2005,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Telomeres shorten during DNA replication; extensive erosion of telomeres likely promotes replicative senescence and chromosomal instability. Telomere length in individual cells has been quantified by flow cytometric analysis of fluorescence in situ hybridization (flow\u2010FISH). To determine the rate of telomere attrition (telomere erosion per cell division), we combined flow\u2010FISH with dye dilution and DNA staining (flow\u2010FISH\u2010DDD) and measured telomere\u2010specific fluorescence in proliferating cells identified by cell generation and cell cycle phase.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Peripheral blood mononuclear cells (PBMC) were stained with the cell division tracking dye carboxyfluorescein diacetate succinimidyl ester (CFSE), stimulated with phytohemagglutinin (PHA), grown for 5\u20136 days, hybridized with a telomere sequence\u2010specific peptide nucleic acid fluorescent probe (PNA\u2010Cy5), counterstained with DAPI, and analyzed by flow cytometry. The cell cycle distribution and cell division generations were respectively identified by analysis of DAPI emission and deconvolution of CFSE emission, and Cy5 emission was used to determine telomere\u2010specific fluorescence, an indicator of telomere length, in each cell.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>In stimulated PBMC, in each cell cycle phase, the telomere\u2010specific fluorescence diminished with increasing cell generation. The rate of decline of the telomere\u2010specific fluorescence per cell generation did not significantly differ between cell cycle phases.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Application of flow\u2010FISH\u2010DDD to measure mean telomere length and the rate of telomere attrition in proliferating cells may find use in studies of ageing and disease, the effects of telomere\u2010modifying agents, and variability between individuals. \u00a9 2005 International Society for Analytical Cytology<\/jats:p><\/jats:sec>","DOI":"10.1002\/cyto.a.20181","type":"journal-article","created":{"date-parts":[[2005,9,14]],"date-time":"2005-09-14T18:26:38Z","timestamp":1126722398000},"page":"53-58","source":"Crossref","is-referenced-by-count":12,"title":["Flow cytometric analysis of fluorescence in situ hybridization with dye dilution and DNA staining (flow\u2010FISH\u2010DDD) to determine telomere length dynamics in proliferating cells"],"prefix":"10.1002","volume":"68A","author":[{"given":"Alan J.","family":"Potter","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark H.","family":"Wener","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,10,19]]},"reference":[{"key":"e_1_2_5_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/nri890"},{"key":"e_1_2_5_3_2","doi-asserted-by":"publisher","DOI":"10.1097\/00001622-200401000-00007"},{"key":"e_1_2_5_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0959-8049(97)00062-2"},{"key":"e_1_2_5_5_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.190.2.157"},{"key":"e_1_2_5_6_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.165.3.1191"},{"key":"e_1_2_5_7_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0609.2004.00305.x"},{"key":"e_1_2_5_8_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V95.6.1883"},{"key":"e_1_2_5_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(03)12384-7"},{"key":"e_1_2_5_10_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/26.16.3651"},{"key":"e_1_2_5_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0898-743"},{"key":"e_1_2_5_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0091-679X(04)75031-1"},{"key":"e_1_2_5_13_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V98.3.597"},{"key":"e_1_2_5_14_2","volume-title":"Current Protocols in Cytometry","author":"Lyons B","year":"2004"},{"key":"e_1_2_5_15_2","volume-title":"Clinical Flow Cytometry: Principles and Application","author":"Rabinovitch PS","year":"1993"},{"key":"e_1_2_5_16_2","doi-asserted-by":"publisher","DOI":"10.1002\/1521-4141(200007)30:7<1894::AID-IMMU1894>3.0.CO;2-N"},{"key":"e_1_2_5_17_2","doi-asserted-by":"publisher","DOI":"10.1006\/excr.2001.5391"},{"key":"e_1_2_5_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0968-0004(02)02110-2"},{"key":"e_1_2_5_19_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.2004.05006.x"},{"key":"e_1_2_5_20_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.97.16.9203"}],"container-title":["Cytometry Part A"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fcyto.a.20181","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.a.20181","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,10]],"date-time":"2023-10-10T22:38:39Z","timestamp":1696977519000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cyto.a.20181"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2005,10,19]]},"references-count":19,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2005,11]]}},"alternative-id":["10.1002\/cyto.a.20181"],"URL":"https:\/\/doi.org\/10.1002\/cyto.a.20181","archive":["Portico"],"relation":{},"ISSN":["1552-4922","1552-4930"],"issn-type":[{"value":"1552-4922","type":"print"},{"value":"1552-4930","type":"electronic"}],"subject":[],"published":{"date-parts":[[2005,10,19]]}}}