{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T15:26:50Z","timestamp":1778254010634,"version":"3.51.4"},"reference-count":53,"publisher":"Elsevier BV","license":[{"start":{"date-parts":[[2023,4,1]],"date-time":"2023-04-01T00:00:00Z","timestamp":1680307200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.elsevier.com\/tdm\/userlicense\/1.0\/"},{"start":{"date-parts":[[2023,4,1]],"date-time":"2023-04-01T00:00:00Z","timestamp":1680307200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.elsevier.com\/legal\/tdmrep-license"},{"start":{"date-parts":[[2023,4,1]],"date-time":"2023-04-01T00:00:00Z","timestamp":1680307200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-017"},{"start":{"date-parts":[[2023,4,1]],"date-time":"2023-04-01T00:00:00Z","timestamp":1680307200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2023,4,1]],"date-time":"2023-04-01T00:00:00Z","timestamp":1680307200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-012"},{"start":{"date-parts":[[2023,4,1]],"date-time":"2023-04-01T00:00:00Z","timestamp":1680307200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2023,4,1]],"date-time":"2023-04-01T00:00:00Z","timestamp":1680307200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-004"}],"funder":[{"DOI":"10.13039\/501100001807","name":"FAPESP","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["elsevier.com","sciencedirect.com"],"crossmark-restriction":true},"short-container-title":["Reproductive Toxicology"],"published-print":{"date-parts":[[2023,4]]},"DOI":"10.1016\/j.reprotox.2023.108353","type":"journal-article","created":{"date-parts":[[2023,2,19]],"date-time":"2023-02-19T22:13:08Z","timestamp":1676844788000},"page":"108353","update-policy":"https:\/\/doi.org\/10.1016\/elsevier_cm_policy","source":"Crossref","is-referenced-by-count":8,"special_numbering":"C","title":["Multiple flow cytometry analysis for assessing human sperm functional characteristics"],"prefix":"10.1016","volume":"117","author":[{"given":"Josiane","family":"de Lima Rosa","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Camila","family":"de Paula Freitas Dell\u2019Aqua","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fabiana Ferreira","family":"de Souza","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gabriela","family":"Missassi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wilma De Grava","family":"Kempinas","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"78","reference":[{"key":"10.1016\/j.reprotox.2023.108353_bib1","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1038\/s41574-021-00598-8","article-title":"Environmental factors in declining human fertility","volume":"18","author":"Skakkeb\u00e6k","year":"2022","journal-title":"Nat. Rev. Endocrionol."},{"key":"10.1016\/j.reprotox.2023.108353_bib2","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1055\/s-0037-1603569","article-title":"Environmental contaminants affecting fertility and somatic health","volume":"35","author":"Chiang","year":"2017","journal-title":"Semin. Reprod. Med."},{"key":"10.1016\/j.reprotox.2023.108353_bib3","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1093\/humupd\/dmx022","article-title":"Temporal trends in sperm count: a systematic review and meta-regression analysis","volume":"23","author":"Levine","year":"2017","journal-title":"Hum. Reprod. Update"},{"key":"10.1016\/j.reprotox.2023.108353_bib4","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1152\/physrev.00017.2015","article-title":"Male reproductive disorders and fertility trends: influence of environment and genetic susceptibility","volume":"96","author":"Skakkeb\u00e6k","year":"2016","journal-title":"Physiol. Rev."},{"key":"10.1016\/j.reprotox.2023.108353_bib5","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1093\/humrep\/deh040","article-title":"Human sperm subpopulations: relationship between functional quality and protein tyrosine phosphorylation","volume":"19","author":"Buffone","year":"2004","journal-title":"Hum. Reprod."},{"issue":"3","key":"10.1016\/j.reprotox.2023.108353_bib6","article-title":"Not all sperm are equal: functional mitochondria characterize a subpopulation of human sperm with better fertilization potential","volume":"6","author":"Souza","year":"2011","journal-title":"Plos One"},{"key":"10.1016\/j.reprotox.2023.108353_bib7","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1002\/ca.22655","article-title":"Comprehensive review of the anatomy and physiology of male ejaculation: premature ejaculation is not a disease","volume":"29","author":"Puppo","year":"2016","journal-title":"Clin. Anat."},{"key":"10.1016\/j.reprotox.2023.108353_bib8","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1111\/rda.13043","article-title":"Flow cytometry in spermatology: a bright future ahead","volume":"52","author":"Ortega-Ferrusola","year":"2017","journal-title":"Reprod. Domest. Anim."},{"key":"10.1016\/j.reprotox.2023.108353_bib9","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1530\/jrf.0.0730441","article-title":"Relationship between the movement characteristics of human spermatozoa and their ability to penetrate cervical mucus and zona-free hamster oocytes","volume":"73","author":"Aitken","year":"1985","journal-title":"J. Reprod. Fertil."},{"key":"10.1016\/j.reprotox.2023.108353_bib10","unstructured":"World Health Organization, WHO laboratory manual for the examination and processing of human semen, fifth ed., Cambridge University Press, Cambridge, 2021."},{"key":"10.1016\/j.reprotox.2023.108353_bib11","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1016\/j.fertnstert.2005.08.024","article-title":"Significance of sperm characteristics in the evaluation of male infertility","volume":"85","author":"Nallella","year":"2006","journal-title":"Fertil. Steril."},{"key":"10.1016\/j.reprotox.2023.108353_bib12","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1590\/S0102-09352010000300006","article-title":"Simultaneous assessment of plasmatic, acrosomal, and mitochondrial membranes in ram sperm by fluorescent probes","volume":"62","author":"Celeghini","year":"2010","journal-title":"Arq. Bras. Med. Vet. Zootec."},{"key":"10.1016\/j.reprotox.2023.108353_bib13","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1016\/j.theriogenology.2004.09.024","article-title":"Flow cytometric evaluation of sperm parameters in relation to fertility potential","volume":"63","author":"Gillan","year":"2005","journal-title":"Theriogenology"},{"key":"10.1016\/j.reprotox.2023.108353_bib14","article-title":"Use of fluorescent dyes for readily recognizing sperm damage","volume":"14","author":"Farah","year":"2013","journal-title":"J. Reprod. Infertil."},{"key":"10.1016\/j.reprotox.2023.108353_bib15","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1016\/j.urology.2014.04.043","article-title":"Functional sperm testing and the role of proteomics in the evaluation of male infertility","volume":"84","author":"Barazani","year":"2014","journal-title":"Urology"},{"key":"10.1016\/j.reprotox.2023.108353_bib16","doi-asserted-by":"crossref","first-page":"163","DOI":"10.3109\/07388551.2015.1128876","article-title":"Flow cytometry: basic principles and applications","volume":"37","author":"Adan","year":"2017","journal-title":"Crit. Rev. Biotechnol."},{"key":"10.1016\/j.reprotox.2023.108353_bib17","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1016\/S0378-4320(01)00160-9","article-title":"Assessment of sperm quality: a flow cytometric approach","volume":"68","author":"Graham","year":"2001","journal-title":"Anim. Reprod. Sci."},{"key":"10.1016\/j.reprotox.2023.108353_bib18","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1080\/gye.16.6.505.521","article-title":"Flow cytometry in human reproductive biology","volume":"16","author":"Garrido","year":"2002","journal-title":"Gynecol. Endocrinol."},{"key":"10.1016\/j.reprotox.2023.108353_bib19","first-page":"351","article-title":"Multiparametric flow cytometry: a relevant tool for sperm function evaluation","volume":"12","author":"Pe\u00f1a","year":"2015","journal-title":"Anim. Reprod."},{"key":"10.1016\/j.reprotox.2023.108353_bib20","doi-asserted-by":"crossref","unstructured":"E. Cordelli, P. Eleuteri, G. Leter, M. Rescia, M. Span\u00f2, Flow cytometry applications in the evaluation of sperm quality: semen analysis, sperm function and DNA integrity, Contraception 72 (20015) 273\u2013279, doi.org\/10.1016\/j.contraception.2005.03.004.","DOI":"10.1016\/j.contraception.2005.03.004"},{"key":"10.1016\/j.reprotox.2023.108353_bib21","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/S0272-0590(82)80059-6","article-title":"Use of animal models for detecting specific alterations in reproduction","volume":"2","author":"Amann","year":"1982","journal-title":"Fundam. Appl. Toxicol."},{"key":"10.1016\/j.reprotox.2023.108353_bib22","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.anireprosci.2007.11.021","article-title":"Saga of a sperm fertility biomarker","volume":"105","author":"Klinefelter","year":"2008","journal-title":"Anim. Reprod. Sci."},{"key":"10.1016\/j.reprotox.2023.108353_bib23","doi-asserted-by":"crossref","first-page":"1171","DOI":"10.1095\/biolreprod38.5.1171","article-title":"Capacitation of bovine sperm by heparin","volume":"38","author":"Parrish","year":"1988","journal-title":"Biol. Reprod."},{"key":"10.1016\/j.reprotox.2023.108353_bib24","first-page":"941","article-title":"Flow cytometric analysis of fertile and subfertile frozen stallion spermatozoa","volume":"9","author":"Freitas-Dell\u2019Aqua","year":"2012","journal-title":"Anim. Reprod."},{"key":"10.1016\/j.reprotox.2023.108353_bib25","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/j.jevs.2018.05.030","article-title":"Superoxide anion is reduced in gradient selected cryopreserved stallion semen despite high mitochondrial potential","volume":"66","author":"Freitas-Dell\u2019Aqua","year":"2018","journal-title":"J. Equine Vet. Sci."},{"key":"10.1016\/j.reprotox.2023.108353_bib26","doi-asserted-by":"crossref","DOI":"10.1016\/j.reprotox.2023.108343","article-title":"SP22 sperm protein as a potential fertility biomarker in humans: a preliminary study","volume":"117","author":"Rosa","year":"2023","journal-title":"Rep. Toxicol."},{"key":"10.1016\/j.reprotox.2023.108353_bib27","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1095\/biolreprod43.1.55","article-title":"Analysis of sperm cell viability, acrosomal integrity, and mitochondrial function using flow cytometry","volume":"43","author":"Graham","year":"1990","journal-title":"Biol. Reprod."},{"key":"10.1016\/j.reprotox.2023.108353_bib28","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/S0378-4320(96)01599-0","article-title":"Simultaneous detection of the acrosomal status and viability of incubated ram spermatozoa using fluorescent markers","volume":"46","author":"Sukardi","year":"1997","journal-title":"Anim. Reprod. Sci."},{"key":"10.1016\/j.reprotox.2023.108353_bib29","doi-asserted-by":"crossref","DOI":"10.1111\/j.1439-0531.2006.00751.x","article-title":"Fluorescent stain method for the simultaneous determination of mitochondrial potential and integrity of plasma and acrosomal membranes in boar sperm","volume":"42","author":"De Andrade","year":"2007","journal-title":"Reprod. Dom. Anim."},{"key":"10.1016\/j.reprotox.2023.108353_bib30","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1371\/journal.pone.0160988","article-title":"Novel flow cytometry analyses of boar sperm viability: can the addition of whole sperm-rich fraction seminal plasma to forzen-thawed boar sperm affect it?","volume":"11","author":"Torres","year":"2016","journal-title":"Plos One"},{"key":"10.1016\/j.reprotox.2023.108353_bib31","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1111\/j.1439-0531.2010.01622.x","article-title":"Probes and techniques for sperm evaluation by flow cytometry","volume":"45","author":"Mart\u00ednez-Pastor","year":"2010","journal-title":"Reprod. Domest. Anim."},{"key":"10.1016\/j.reprotox.2023.108353_bib32","unstructured":"D.A. McCarthy, Fluorochromes and fluorescence. Humana Press Inc, New Jersey, 2007."},{"key":"10.1016\/j.reprotox.2023.108353_bib33","series-title":"Detection of Hematopoietic Stem Cells by Flow Cytometry","author":"Lin","year":"2011"},{"key":"10.1016\/j.reprotox.2023.108353_bib34","doi-asserted-by":"crossref","first-page":"1073","DOI":"10.1016\/j.theriogenology.2009.06.027","article-title":"Washing increases the susceptibility to exogenous oxidative stress in red deer spermatozoa","volume":"72","author":"Dom\u00ednguez-Rebolledo","year":"2009","journal-title":"Theriogenology"},{"key":"10.1016\/j.reprotox.2023.108353_bib35","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1080\/014850190924485","article-title":"Single UV excitation of hoechst 33342 and propidium iodide for viability assessment of rhesus monkey spermatozoa using flow cytometry","volume":"51","author":"Cai","year":"2005","journal-title":"Arch. Androl."},{"key":"10.1016\/j.reprotox.2023.108353_bib36","first-page":"1","article-title":"Optimized stem cell detection using the dyecycle-triggered side population phenotype","author":"Boesch","year":"2016","journal-title":"Stem Cells Int"},{"key":"10.1016\/j.reprotox.2023.108353_bib37","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1038\/aja.2011.15","article-title":"Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art","volume":"13","author":"Hossain","year":"2011","journal-title":"Asian J. Androl."},{"key":"10.1016\/j.reprotox.2023.108353_bib38","series-title":"Physiology of Male Gametogenesis","author":"Sharma","year":"2007"},{"key":"10.1016\/j.reprotox.2023.108353_bib39","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1006\/abbi.2000.1880","article-title":"Mammalian sperm acrosome: formation, contents, and function","volume":"379","author":"Abou-Haila","year":"2000","journal-title":"Arch. Biochem. Biophys."},{"key":"10.1016\/j.reprotox.2023.108353_bib40","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1095\/biolreprod53.2.276","article-title":"Viability assessment of mammalian sperm using SYBR-14 and propidium iodide","volume":"53","author":"Garner","year":"1995","journal-title":"Biol. Reprod."},{"key":"10.1016\/j.reprotox.2023.108353_bib41","doi-asserted-by":"crossref","DOI":"10.1038\/nprot.2006.238","article-title":"Analysis of apoptosis by propidium iodide staining and flow cytometry","volume":"1","author":"Riccardi","year":"2006","journal-title":"Nat. Protoc."},{"key":"10.1016\/j.reprotox.2023.108353_bib42","doi-asserted-by":"crossref","DOI":"10.1016\/S0015-0282(03)00983-X","article-title":"Alterations in mitochondria membrane potential and oxidative stress in infertile men: a prospective observational study","volume":"80","author":"Wang","year":"2003","journal-title":"Fertil. Steril."},{"key":"10.1016\/j.reprotox.2023.108353_bib43","doi-asserted-by":"crossref","DOI":"10.1007\/s00204-013-1034-4","article-title":"Oxidative stress: the mitochondrial-dependent and mitochondria-independent pathways of apoptosis","volume":"87","author":"Sinha","year":"2013","journal-title":"Arch. Toxicol."},{"key":"10.1016\/j.reprotox.2023.108353_bib44","doi-asserted-by":"crossref","DOI":"10.1111\/j.2047-2927.2013.00098.x","article-title":"UVB irradiation as a tool to assess ROS-induced damage in human spermatozoa","volume":"1","author":"Amaral","year":"2013","journal-title":"Andrology"},{"key":"10.1016\/j.reprotox.2023.108353_bib45","doi-asserted-by":"crossref","DOI":"10.1530\/REP-08-0357","article-title":"Reactive oxygen species generators affect quality parameters and apoptosis markers differently in red deer spermatozoa","volume":"137","author":"Mart\u00ednez-Pastor","year":"2009","journal-title":"Reproduction"},{"key":"10.1016\/j.reprotox.2023.108353_bib46","doi-asserted-by":"crossref","DOI":"10.1016\/j.fertnstert.2008.10.068","article-title":"Sperm viability, apoptosis, and intracellular reactive oxygen species levels in human spermatozoa before and after induction of oxidative stress","volume":"93","author":"Mahfouz","year":"2010","journal-title":"Fertil. Steril."},{"key":"10.1016\/j.reprotox.2023.108353_bib47","doi-asserted-by":"crossref","DOI":"10.1016\/j.anireprosci.2012.01.003","article-title":"Effect of Hoechst 33342 on stallion spermatozoa incubated in KMT or Tyrodes modified INRA96","volume":"131","author":"Balao da Silva","year":"2012","journal-title":"Anim. Reprod. Sci."},{"key":"10.1016\/j.reprotox.2023.108353_bib48","article-title":"Sperm membrane integrity and stability after selection of cryopreserved ovine semen on colloidal solutions","volume":"50","author":"Bergstein-Galan","year":"2017","journal-title":"Andrologia"},{"key":"10.1016\/j.reprotox.2023.108353_bib49","doi-asserted-by":"crossref","DOI":"10.1071\/RD08002","article-title":"Mitochondrial activity and forward scatter vary in necrotic, apoptotic and membrane-intact spermatozoa subpopulations","volume":"20","author":"Mart\u00ednez-Pastor","year":"2008","journal-title":"Reprod. Fertil. Dev."},{"key":"10.1016\/j.reprotox.2023.108353_bib50","unstructured":"P. Allen, D. Davies, Apoptosis detection by flow cytometry. Humana Press Inc, New Jersey, 2007."},{"key":"10.1016\/j.reprotox.2023.108353_bib51","doi-asserted-by":"crossref","DOI":"10.1016\/j.anireprosci.2018.02.020","article-title":"Effects of coenzyme Q10 on semen cryopreservation of stallions classified as having good or bad semen freezing ability","volume":"192","author":"Carneiro","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"10.1016\/j.reprotox.2023.108353_bib52","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1111\/j.2047-2927.2012.00056.x","article-title":"On methods for the detection of reactive oxygen species generation by human spermatozoa: analysis of the cellular responses to catechol oestrogen, lipid aldehyde, menadione and arachidonic acid","volume":"1","author":"Aitken","year":"2013","journal-title":"Andrology"},{"key":"10.1016\/j.reprotox.2023.108353_bib53","doi-asserted-by":"crossref","DOI":"10.1016\/j.freeradbiomed.2015.05.032","article-title":"Low micromolar concentrations of the superoxide probe MitoSOX uncouple neural mitochondria and inhibit complex IV","volume":"86","author":"Roelofs","year":"2015","journal-title":"Free Radic. Biol. Med."}],"container-title":["Reproductive Toxicology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.elsevier.com\/content\/article\/PII:S0890623823000266?httpAccept=text\/xml","content-type":"text\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.elsevier.com\/content\/article\/PII:S0890623823000266?httpAccept=text\/plain","content-type":"text\/plain","content-version":"vor","intended-application":"text-mining"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:59:52Z","timestamp":1760191192000},"score":1,"resource":{"primary":{"URL":"https:\/\/linkinghub.elsevier.com\/retrieve\/pii\/S0890623823000266"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4]]},"references-count":53,"alternative-id":["S0890623823000266"],"URL":"https:\/\/doi.org\/10.1016\/j.reprotox.2023.108353","relation":{},"ISSN":["0890-6238"],"issn-type":[{"value":"0890-6238","type":"print"}],"subject":[],"published":{"date-parts":[[2023,4]]},"assertion":[{"value":"Elsevier","name":"publisher","label":"This article is maintained by"},{"value":"Multiple flow cytometry analysis for assessing human sperm functional characteristics","name":"articletitle","label":"Article Title"},{"value":"Reproductive Toxicology","name":"journaltitle","label":"Journal Title"},{"value":"https:\/\/doi.org\/10.1016\/j.reprotox.2023.108353","name":"articlelink","label":"CrossRef DOI link to publisher maintained version"},{"value":"article","name":"content_type","label":"Content Type"},{"value":"\u00a9 2023 Elsevier Inc. All rights reserved.","name":"copyright","label":"Copyright"}],"article-number":"108353"}}