Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata

Detalhes bibliográficos
Autor(a) principal: Brianezi, Gabrielli [UNESP]
Data de Publicação: 2009
Outros Autores: De Camargo, João Lauro Viana [UNESP], Miot, Hélio Amante [UNESP]
Tipo de documento: Artigo
Idioma: por
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://hdl.handle.net/11449/225724
Resumo: Background: Comet assay or single cell gel electrophoresis is a useful and widely applied technique for the assessment of DNA damage and repair in individual cells. Nuclei can be stained with fluorescence methods or silver salts. The latter has technical advantages such as the type of microscope used and the possibility of slide storage. Comet analysis may be performed visually, however, there is the disadvantage of subjective results, which can be minimized by automated digital analysis. Objectives: Development and validation of digital analysis method for silver stained comet assays. Methods: Fifty comets were photographed in a standardized way and printed on paper. Before and after being automatically preprocessed by Imagej 1.38x software, the images were manually measured and classified into five categories by three evaluators. Their estimates were compared as to their correlation and reproducibility. Afterwards, an algorithm for automated digital analysis of the comet measurements was developed based on statistical filters of median and minimum. These results were compared with those manually and visually estimated after preprocessing. Results: Manual measurements of preprocessed images showed higher intraclass correlation than the original ones. Automated results had high correlation with the pre-processed manual measurements, suggesting that this system increases objectivity and can be used in the estimation of comet parameters. Conclusions: Digital analysis of silver stained comet assay proved to be feasible and better reproducible than the visual analysis.
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spelling Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prataDevelopment and validation of a quantitative image analysis method to evaluate comet assay (silver staining)Comet assayGenotoxicityImage analysisQuantitative methodBackground: Comet assay or single cell gel electrophoresis is a useful and widely applied technique for the assessment of DNA damage and repair in individual cells. Nuclei can be stained with fluorescence methods or silver salts. The latter has technical advantages such as the type of microscope used and the possibility of slide storage. Comet analysis may be performed visually, however, there is the disadvantage of subjective results, which can be minimized by automated digital analysis. Objectives: Development and validation of digital analysis method for silver stained comet assays. Methods: Fifty comets were photographed in a standardized way and printed on paper. Before and after being automatically preprocessed by Imagej 1.38x software, the images were manually measured and classified into five categories by three evaluators. Their estimates were compared as to their correlation and reproducibility. Afterwards, an algorithm for automated digital analysis of the comet measurements was developed based on statistical filters of median and minimum. These results were compared with those manually and visually estimated after preprocessing. Results: Manual measurements of preprocessed images showed higher intraclass correlation than the original ones. Automated results had high correlation with the pre-processed manual measurements, suggesting that this system increases objectivity and can be used in the estimation of comet parameters. Conclusions: Digital analysis of silver stained comet assay proved to be feasible and better reproducible than the visual analysis.Núcleo de Avaliação do Impacto Ambiental Sobre A Saúde Humana (TOXICAM) Departamento de Patologia Universidade Estadual Paulista (FMB/UNESP)Departamento de Patologia; Responsável Pelo TOXICAM Departamento de Patologia da FMB/UNESPDepartamento de Dermatologia e Radioterapia da FMB/UNESP Campus Universitário de Rublão Júnior, CEP. 18618-000-Botucatu-SPNúcleo de Avaliação do Impacto Ambiental Sobre A Saúde Humana (TOXICAM) Departamento de Patologia Universidade Estadual Paulista (FMB/UNESP)Departamento de Patologia; Responsável Pelo TOXICAM Departamento de Patologia da FMB/UNESPDepartamento de Dermatologia e Radioterapia da FMB/UNESP Campus Universitário de Rublão Júnior, CEP. 18618-000-Botucatu-SPUniversidade Estadual Paulista (UNESP)Brianezi, Gabrielli [UNESP]De Camargo, João Lauro Viana [UNESP]Miot, Hélio Amante [UNESP]2022-04-28T20:59:05Z2022-04-28T20:59:05Z2009-08-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article325-334Jornal Brasileiro de Patologia e Medicina Laboratorial, v. 45, n. 4, p. 325-334, 2009.1676-24441678-4774http://hdl.handle.net/11449/2257242-s2.0-73449104577Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPporJornal Brasileiro de Patologia e Medicina Laboratorialinfo:eu-repo/semantics/openAccess2024-09-03T13:14:31Zoai:repositorio.unesp.br:11449/225724Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestrepositoriounesp@unesp.bropendoar:29462024-09-03T13:14:31Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata
Development and validation of a quantitative image analysis method to evaluate comet assay (silver staining)
title Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata
spellingShingle Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata
Brianezi, Gabrielli [UNESP]
Comet assay
Genotoxicity
Image analysis
Quantitative method
title_short Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata
title_full Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata
title_fullStr Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata
title_full_unstemmed Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata
title_sort Desenvolvimento e validação de técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata
author Brianezi, Gabrielli [UNESP]
author_facet Brianezi, Gabrielli [UNESP]
De Camargo, João Lauro Viana [UNESP]
Miot, Hélio Amante [UNESP]
author_role author
author2 De Camargo, João Lauro Viana [UNESP]
Miot, Hélio Amante [UNESP]
author2_role author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (UNESP)
dc.contributor.author.fl_str_mv Brianezi, Gabrielli [UNESP]
De Camargo, João Lauro Viana [UNESP]
Miot, Hélio Amante [UNESP]
dc.subject.por.fl_str_mv Comet assay
Genotoxicity
Image analysis
Quantitative method
topic Comet assay
Genotoxicity
Image analysis
Quantitative method
description Background: Comet assay or single cell gel electrophoresis is a useful and widely applied technique for the assessment of DNA damage and repair in individual cells. Nuclei can be stained with fluorescence methods or silver salts. The latter has technical advantages such as the type of microscope used and the possibility of slide storage. Comet analysis may be performed visually, however, there is the disadvantage of subjective results, which can be minimized by automated digital analysis. Objectives: Development and validation of digital analysis method for silver stained comet assays. Methods: Fifty comets were photographed in a standardized way and printed on paper. Before and after being automatically preprocessed by Imagej 1.38x software, the images were manually measured and classified into five categories by three evaluators. Their estimates were compared as to their correlation and reproducibility. Afterwards, an algorithm for automated digital analysis of the comet measurements was developed based on statistical filters of median and minimum. These results were compared with those manually and visually estimated after preprocessing. Results: Manual measurements of preprocessed images showed higher intraclass correlation than the original ones. Automated results had high correlation with the pre-processed manual measurements, suggesting that this system increases objectivity and can be used in the estimation of comet parameters. Conclusions: Digital analysis of silver stained comet assay proved to be feasible and better reproducible than the visual analysis.
publishDate 2009
dc.date.none.fl_str_mv 2009-08-01
2022-04-28T20:59:05Z
2022-04-28T20:59:05Z
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 Jornal Brasileiro de Patologia e Medicina Laboratorial, v. 45, n. 4, p. 325-334, 2009.
1676-2444
1678-4774
http://hdl.handle.net/11449/225724
2-s2.0-73449104577
identifier_str_mv Jornal Brasileiro de Patologia e Medicina Laboratorial, v. 45, n. 4, p. 325-334, 2009.
1676-2444
1678-4774
2-s2.0-73449104577
url http://hdl.handle.net/11449/225724
dc.language.iso.fl_str_mv por
language por
dc.relation.none.fl_str_mv Jornal Brasileiro de Patologia e Medicina Laboratorial
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 325-334
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv repositoriounesp@unesp.br
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