Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status
Main Author: | |
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Publication Date: | 2009 |
Other Authors: | , , , , , |
Format: | Article |
Language: | eng |
Source: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Download full: | http://hdl.handle.net/10316/12311 https://doi.org/10.1093/humrep/den365 |
Summary: | BACKGROUND: Sperm chromatin status and nuclear DNA damage can be detected using well-established assays. However, most techniques are time-consuming and/or involve elaborate protocols and equipment. We have recently developed a simple and fast method to monitor sperm chromatin status in field conditions using the Diff-Quik assay which is employed in fertility clinics to assess sperm morphology with standard bright field microscopy. In the present study, we demonstrate that any Diff-Quik-like stain can easily, reproducibly and routinely monitor human sperm chromatin status as well. METHODS: Different Diff-Quik-like stains were used to assess sperm morphology and the presence of abnormal dark nuclear staining in human sperm from four ART centres. The TUNEL assay was performed in the same samples, and fertility outcomes were assessed. RESULTS: A significant correlation was found between TUNEL-positive sperm and dark sperm nuclei. Moreover, associations were also found between the percentage of dark sperm nuclei and seminal parameters, embryo development rate, embryo quality and clinical pregnancy, as well as with cryptorchidism, and there was a tendency towards an association with age. A value of 32% abnormal staining is suggested as a predictive threshold for embryo development and pregnancy. CONCLUSIONS: Our results show that any Diff-Quik-like stain, already implemented in most laboratories to assess sperm morphology, can be adapted as an indicator for chromatin status in human sperm |
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Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin statusHuman spermChromatinNuclear DNADiff-Quik stainingSperm morphologyBACKGROUND: Sperm chromatin status and nuclear DNA damage can be detected using well-established assays. However, most techniques are time-consuming and/or involve elaborate protocols and equipment. We have recently developed a simple and fast method to monitor sperm chromatin status in field conditions using the Diff-Quik assay which is employed in fertility clinics to assess sperm morphology with standard bright field microscopy. In the present study, we demonstrate that any Diff-Quik-like stain can easily, reproducibly and routinely monitor human sperm chromatin status as well. METHODS: Different Diff-Quik-like stains were used to assess sperm morphology and the presence of abnormal dark nuclear staining in human sperm from four ART centres. The TUNEL assay was performed in the same samples, and fertility outcomes were assessed. RESULTS: A significant correlation was found between TUNEL-positive sperm and dark sperm nuclei. Moreover, associations were also found between the percentage of dark sperm nuclei and seminal parameters, embryo development rate, embryo quality and clinical pregnancy, as well as with cryptorchidism, and there was a tendency towards an association with age. A value of 32% abnormal staining is suggested as a predictive threshold for embryo development and pregnancy. CONCLUSIONS: Our results show that any Diff-Quik-like stain, already implemented in most laboratories to assess sperm morphology, can be adapted as an indicator for chromatin status in human spermOxford University Press2009info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/12311http://hdl.handle.net/10316/12311https://doi.org/10.1093/humrep/den365engHuman Reproduction. 24:1 (2009) 28-360268-1161Sousa, Ana Paula M.Tavares, Renata S.Velez de la Calle, Juan FelipeFigueiredo, HelenaAlmeida, VascoAlmeida-Santos, TeresaRamalho-Santos, Joãoinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2021-11-02T09:49:16Zoai:estudogeral.uc.pt:10316/12311Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:55:40.341587Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status |
title |
Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status |
spellingShingle |
Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status Sousa, Ana Paula M. Human sperm Chromatin Nuclear DNA Diff-Quik staining Sperm morphology |
title_short |
Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status |
title_full |
Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status |
title_fullStr |
Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status |
title_full_unstemmed |
Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status |
title_sort |
Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status |
author |
Sousa, Ana Paula M. |
author_facet |
Sousa, Ana Paula M. Tavares, Renata S. Velez de la Calle, Juan Felipe Figueiredo, Helena Almeida, Vasco Almeida-Santos, Teresa Ramalho-Santos, João |
author_role |
author |
author2 |
Tavares, Renata S. Velez de la Calle, Juan Felipe Figueiredo, Helena Almeida, Vasco Almeida-Santos, Teresa Ramalho-Santos, João |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Sousa, Ana Paula M. Tavares, Renata S. Velez de la Calle, Juan Felipe Figueiredo, Helena Almeida, Vasco Almeida-Santos, Teresa Ramalho-Santos, João |
dc.subject.por.fl_str_mv |
Human sperm Chromatin Nuclear DNA Diff-Quik staining Sperm morphology |
topic |
Human sperm Chromatin Nuclear DNA Diff-Quik staining Sperm morphology |
description |
BACKGROUND: Sperm chromatin status and nuclear DNA damage can be detected using well-established assays. However, most techniques are time-consuming and/or involve elaborate protocols and equipment. We have recently developed a simple and fast method to monitor sperm chromatin status in field conditions using the Diff-Quik assay which is employed in fertility clinics to assess sperm morphology with standard bright field microscopy. In the present study, we demonstrate that any Diff-Quik-like stain can easily, reproducibly and routinely monitor human sperm chromatin status as well. METHODS: Different Diff-Quik-like stains were used to assess sperm morphology and the presence of abnormal dark nuclear staining in human sperm from four ART centres. The TUNEL assay was performed in the same samples, and fertility outcomes were assessed. RESULTS: A significant correlation was found between TUNEL-positive sperm and dark sperm nuclei. Moreover, associations were also found between the percentage of dark sperm nuclei and seminal parameters, embryo development rate, embryo quality and clinical pregnancy, as well as with cryptorchidism, and there was a tendency towards an association with age. A value of 32% abnormal staining is suggested as a predictive threshold for embryo development and pregnancy. CONCLUSIONS: Our results show that any Diff-Quik-like stain, already implemented in most laboratories to assess sperm morphology, can be adapted as an indicator for chromatin status in human sperm |
publishDate |
2009 |
dc.date.none.fl_str_mv |
2009 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10316/12311 http://hdl.handle.net/10316/12311 https://doi.org/10.1093/humrep/den365 |
url |
http://hdl.handle.net/10316/12311 https://doi.org/10.1093/humrep/den365 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Human Reproduction. 24:1 (2009) 28-36 0268-1161 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Oxford University Press |
publisher.none.fl_str_mv |
Oxford University Press |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
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1799133843257556992 |