{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,16]],"date-time":"2026-05-16T03:53:51Z","timestamp":1778903631683,"version":"3.51.4"},"reference-count":46,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2017,7,5]],"date-time":"2017-07-05T00:00:00Z","timestamp":1499212800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01ES017457"],"award-info":[{"award-number":["R01ES017457"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["S10OD010710"],"award-info":[{"award-number":["S10OD010710"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["S10RR025565"],"award-info":[{"award-number":["S10RR025565"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2017,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Structural and numerical sperm chromosomal aberrations result from abnormal meiosis and are directly linked to infertility. Any live births that arise from aneuploid conceptuses can result in syndromes such as Kleinfelter, Turners, XYY and Edwards. Multi\u2010probe fluorescence <jats:italic>in situ<\/jats:italic> hybridization (FISH) is commonly used to study sperm aneuploidy, however manual FISH scoring in sperm samples is labor\u2010intensive and introduces errors. Automated scoring methods are continuously evolving. One challenging aspect for optimizing automated sperm FISH scoring has been the overlap in excitation and emission of the fluorescent probes used to enumerate the chromosomes of interest. Our objective was to demonstrate the feasibility of combining confocal microscopy and spectral imaging with high\u2010throughput methods for accurately measuring sperm aneuploidy. Our approach used confocal microscopy to analyze numerical chromosomal abnormalities in human sperm using enhanced slide preparation and rigorous semi\u2010automated scoring methods. FISH for chromosomes X, Y, and 18 was conducted to determine sex chromosome disomy in sperm nuclei. Application of online spectral linear unmixing was used for effective separation of four fluorochromes while decreasing data acquisition time. Semi\u2010automated image processing, segmentation, classification, and scoring were performed on 10 slides using custom image processing and analysis software and results were compared with manual methods. No significant differences in disomy frequencies were seen between the semi automated and manual methods. Samples treated with pepsin were observed to have reduced background autofluorescence and more uniform distribution of cells. These results demonstrate that semi\u2010automated methods using spectral imaging on a confocal platform are a feasible approach for analyzing numerical chromosomal aberrations in sperm, and are comparable to manual methods. \u00a9 2017 International Society for Advancement of Cytometry<\/jats:p>","DOI":"10.1002\/cyto.a.23126","type":"journal-article","created":{"date-parts":[[2017,7,5]],"date-time":"2017-07-05T15:48:37Z","timestamp":1499269717000},"page":"859-866","source":"Crossref","is-referenced-by-count":5,"title":["Semi\u2010automated scoring of triple\u2010probe FISH in human sperm using confocal microscopy"],"prefix":"10.1002","volume":"91","author":[{"given":"Francesca","family":"Branch","sequence":"first","affiliation":[{"name":"Department of Environmental and Occupational Health Milken Institute School of Public Health, George Washington University Washington, DC"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"GiaLinh","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Department of Environmental and Occupational Health Milken Institute School of Public Health, George Washington University Washington, DC"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicholas","family":"Porter","sequence":"additional","affiliation":[{"name":"Department of Environmental and Occupational Health Milken Institute School of Public Health, George Washington University Washington, DC"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heather A.","family":"Young","sequence":"additional","affiliation":[{"name":"Department of Epidemiology and Biostatistics Milken Institute School of Public Health, George Washington University Washington, DC"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sheena E.","family":"Martenies","sequence":"additional","affiliation":[{"name":"Department of Environmental and Occupational Health Milken Institute School of Public Health, George Washington University Washington, DC"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nathan","family":"McCray","sequence":"additional","affiliation":[{"name":"Department of Environmental and Occupational Health Milken Institute School of Public Health, George Washington University Washington, DC"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Glen","family":"Deloid","sequence":"additional","affiliation":[{"name":"Department of Environmental Health Harvard School of Public Health Boston, Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anastas","family":"Popratiloff","sequence":"additional","affiliation":[{"name":"Nanofabrication and Imaging Center George Washington University Washington, DC"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Melissa J.","family":"Perry","sequence":"additional","affiliation":[{"name":"Department of Environmental and Occupational Health Milken Institute School of Public Health, George Washington University Washington, DC"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2017,7,5]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/35066065"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajmg.10391"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1159\/000056773"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1053\/beog.2000.0124"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0074-7696(08)61349-2"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/17.3.576"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1007\/s004390050964"},{"key":"e_1_2_6_9_1","first-page":"267","article-title":"The XYY syndrome: A follow\u2010up study on 38 boys","volume":"14","author":"Geerts M","year":"2003","journal-title":"Genet Couns"},{"key":"e_1_2_6_10_1","first-page":"669","article-title":"Trisomy 18: Studies of the parent and cell division of origin and the effect of aberrant recombination on nondisjunction","volume":"56","author":"Fisher JM","year":"1995","journal-title":"Am J Hum Genet"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1177\/1756287210373758"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/1361-6374(199606)4:2<65::AID-BIO4>3.0.CO;2-D"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1159\/000064038"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.00633"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng1196-312"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1383-5718(00)00141-8"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fertnstert.2007.06.095"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1186\/2051-4190-23-13"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20468"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.21078"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/aja.2009.85"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0320(19970501)28:1<1::AID-CYTO1>3.0.CO;2-K"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fertnstert.2008.07.1779"},{"key":"e_1_2_6_24_1","doi-asserted-by":"crossref","first-page":"926","DOI":"10.1128\/aem.61.3.926-936.1995","article-title":"Fully automatic determination of soil bacterium numbers, cell volumes, and frequencies of dividing cells by confocal laser scanning microscopy and image analysis","volume":"61","author":"Bloem J","year":"1995","journal-title":"Appl Environ Microbiol"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1759(99)00240-9"},{"key":"e_1_2_6_26_1","doi-asserted-by":"crossref","unstructured":"WHO (World Health Organization). Laboratory manual for the examination and processing of human semen.2010;5th edition.","DOI":"10.1038\/aja.2008.57"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.1289\/ehp.1104017"},{"key":"e_1_2_6_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.theriogenology.2006.11.014"},{"key":"e_1_2_6_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fertnstert.2011.04.009"},{"key":"e_1_2_6_30_1","doi-asserted-by":"publisher","DOI":"10.2144\/01316bt01"},{"key":"e_1_2_6_31_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1101134108"},{"key":"e_1_2_6_32_1","doi-asserted-by":"publisher","DOI":"10.1093\/humupd\/dmp048"},{"key":"e_1_2_6_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fertnstert.2007.11.002"},{"key":"e_1_2_6_34_1","doi-asserted-by":"publisher","DOI":"10.1159\/000079575"},{"key":"e_1_2_6_35_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00272374"},{"key":"e_1_2_6_36_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.990140704"},{"key":"e_1_2_6_37_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20315"},{"key":"e_1_2_6_38_1","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.humrep.a138540"},{"key":"e_1_2_6_39_1","doi-asserted-by":"publisher","DOI":"10.1309\/FTT59GWKDWH69FB0"},{"key":"e_1_2_6_40_1","doi-asserted-by":"publisher","DOI":"10.1177\/153303461000900302"},{"key":"e_1_2_6_41_1","doi-asserted-by":"publisher","DOI":"10.1186\/1746-1596-8-17"},{"key":"e_1_2_6_42_1","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6890-13-13"},{"key":"e_1_2_6_43_1","article-title":"Fully automated fluorescent in situ hybridization (FISH) staining and digital analysis of HER2 in breast cancer: A validation study","volume":"10","author":"Logt EMJ","year":"2015","journal-title":"PLoS ONE"},{"key":"e_1_2_6_44_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-60761-847-8_17"},{"key":"e_1_2_6_45_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20319"},{"key":"e_1_2_6_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2006.07.122"},{"key":"e_1_2_6_47_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20292"}],"container-title":["Cytometry Part A"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fcyto.a.23126","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.a.23126","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,12]],"date-time":"2023-09-12T21:16:39Z","timestamp":1694553399000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cyto.a.23126"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,7,5]]},"references-count":46,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2017,9]]}},"alternative-id":["10.1002\/cyto.a.23126"],"URL":"https:\/\/doi.org\/10.1002\/cyto.a.23126","archive":["Portico"],"relation":{},"ISSN":["1552-4922","1552-4930"],"issn-type":[{"value":"1552-4922","type":"print"},{"value":"1552-4930","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,7,5]]}}}