Trypan blue exclusion assay by flow cytometry

Detalhes bibliográficos
Autor(a) principal: Freitas, Bethânia Alves de Avelar
Data de Publicação: 2014
Outros Autores: Almeida, Valéria Gomes de, Pinto, Mauro Cunha Xavier, Mourão, Flávio Afonso Gonçalves, Massensini, Andre Ricardo, Martins Filho, Olindo Assis, Vieira, Etel Rocha, Melo, Gustavo Eustáquio Brito Alvim de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/29669
Resumo: 2020-01-01
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spelling Freitas, Bethânia Alves de AvelarAlmeida, Valéria Gomes dePinto, Mauro Cunha XavierMourão, Flávio Afonso GonçalvesMassensini, Andre RicardoMartins Filho, Olindo AssisVieira, Etel RochaMelo, Gustavo Eustáquio Brito Alvim de2018-10-19T19:16:20Z2018-10-19T19:16:20Z2014FREITAS, Bethânia Alves de Avelar et al. Trypan blue exclusion assay by flow cytometry. Braz J Med Biol Res. , v. 47, n. 4, p. 307-15, 2014.0100-879Xhttps://www.arca.fiocruz.br/handle/icict/2966910.1590/1414-431X20143437engAssociação Brasileira de Divulgação CientíficaAzul tripanoFluorescênciaCitometria de fluxoCitotoxicidadeTrypan blueFluorescenceFlow cytometryCytotoxicityTrypan blue exclusion assay by flow cytometryinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article2020-01-01Universidade Federal dos Vales do Jequitinhonha e Mucuri. Departamento de Farmácia. Laboratório de Imunologia. Programa Multicêntrico de Pós-graduação em Ciências Fisiológicas, Diamantina, MG, Brasil / Universidade Federal dos Vales do Jequitinhonha e Mucuri. Instituto de Ciência e Tecnologia. Diamantina, MG, Brasil.Universidade Federal dos Vales do Jequitinhonha e Mucuri. Departamento de Farmácia. Laboratório de Imunologia. Programa Multicêntrico de Pós-graduação em Ciências Fisiológicas, Diamantina, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Fisiologia e Farmacologia. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Fisiologia e Farmacologia. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Fisiologia e Farmacologia. Belo Horizonte, MG, Brasil.Fundação Osvaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil.Universidade Federal dos Vales do Jequitinhonha e Mucuri. Departamento de Farmácia. Laboratório de Imunologia. Programa Multicêntrico de Pós-graduação em Ciências Fisiológicas, Diamantina, MG, Brasil.Universidade Federal dos Vales do Jequitinhonha e Mucuri. Departamento de Farmácia. Laboratório de Imunologia. Programa Multicêntrico de Pós-graduação em Ciências Fisiológicas, Diamantina, MG, Brasil.Dye exclusion tests are used to determine the number of live and dead cells. These assays are based on the principle that intact plasma membranes in live cells exclude specific dyes, whereas dead cells do not. Although widely used, the trypan blue (TB) exclusion assay has limitations. The dye can be incorporated by live cells after a short exposure time, and personal reliability, related to the expertise of the analyst, can affect the results. We propose an alternative assay for evaluating cell viability that combines the TB exclusion test and the high sensitivity of the flow cytometry technique. Previous studies have demonstrated the ability of TB to emit fluorescence when complexed with proteins. According to our results, TB/bovine serum albumin and TB/cytoplasmic protein complexes emit fluorescence at 660 nm, which is detectable by flow cytometry using a 650-nm low-pass band filter. TB at 0.002% (w/v) was defined as the optimum concentration for distinguishing unstained living cells from fluorescent dead cells, and fluorescence emission was stable for 30 min after cell treatment. Although previous studies have shown that TB promotes green fluorescence quenching, TB at 0.002% did not interfere with green fluorescence in human live T-cells stained with anti-CD3/fluorescein isothiocyanate (FITC) monoclonal antibody. We observed a high correlation between the percentage of propidium iodide+CD3/FITC+ and TB+CD3/FITC+ cells, as well as similar double-stained cell profiles in flow cytometry dot-plot graphs. Taken together, the results indicate that a TB exclusion assay by flow cytometry can be employed as an alternative tool for quick and reliable cell viability analysis.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83082https://www.arca.fiocruz.br/bitstream/icict/29669/1/license.txt9193a7c197bc67acd023525e72a03240MD51ORIGINALTrypan blue exclusion assay by flow cytometry.pdfTrypan blue exclusion assay by flow cytometry.pdfapplication/pdf6516733https://www.arca.fiocruz.br/bitstream/icict/29669/2/Trypan%20blue%20exclusion%20assay%20by%20flow%20cytometry.pdfedbd16c6b878a46860d88bd5e4e58bc3MD52TEXTTrypan blue exclusion assay by flow cytometry.pdf.txtTrypan blue exclusion assay by flow cytometry.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv Trypan blue exclusion assay by flow cytometry
title Trypan blue exclusion assay by flow cytometry
spellingShingle Trypan blue exclusion assay by flow cytometry
Freitas, Bethânia Alves de Avelar
Azul tripano
Fluorescência
Citometria de fluxo
Citotoxicidade
Trypan blue
Fluorescence
Flow cytometry
Cytotoxicity
title_short Trypan blue exclusion assay by flow cytometry
title_full Trypan blue exclusion assay by flow cytometry
title_fullStr Trypan blue exclusion assay by flow cytometry
title_full_unstemmed Trypan blue exclusion assay by flow cytometry
title_sort Trypan blue exclusion assay by flow cytometry
author Freitas, Bethânia Alves de Avelar
author_facet Freitas, Bethânia Alves de Avelar
Almeida, Valéria Gomes de
Pinto, Mauro Cunha Xavier
Mourão, Flávio Afonso Gonçalves
Massensini, Andre Ricardo
Martins Filho, Olindo Assis
Vieira, Etel Rocha
Melo, Gustavo Eustáquio Brito Alvim de
author_role author
author2 Almeida, Valéria Gomes de
Pinto, Mauro Cunha Xavier
Mourão, Flávio Afonso Gonçalves
Massensini, Andre Ricardo
Martins Filho, Olindo Assis
Vieira, Etel Rocha
Melo, Gustavo Eustáquio Brito Alvim de
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Freitas, Bethânia Alves de Avelar
Almeida, Valéria Gomes de
Pinto, Mauro Cunha Xavier
Mourão, Flávio Afonso Gonçalves
Massensini, Andre Ricardo
Martins Filho, Olindo Assis
Vieira, Etel Rocha
Melo, Gustavo Eustáquio Brito Alvim de
dc.subject.other.pt_BR.fl_str_mv Azul tripano
Fluorescência
Citometria de fluxo
Citotoxicidade
topic Azul tripano
Fluorescência
Citometria de fluxo
Citotoxicidade
Trypan blue
Fluorescence
Flow cytometry
Cytotoxicity
dc.subject.en.pt_BR.fl_str_mv Trypan blue
Fluorescence
Flow cytometry
Cytotoxicity
description 2020-01-01
publishDate 2014
dc.date.issued.fl_str_mv 2014
dc.date.accessioned.fl_str_mv 2018-10-19T19:16:20Z
dc.date.available.fl_str_mv 2018-10-19T19:16:20Z
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.citation.fl_str_mv FREITAS, Bethânia Alves de Avelar et al. Trypan blue exclusion assay by flow cytometry. Braz J Med Biol Res. , v. 47, n. 4, p. 307-15, 2014.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/29669
dc.identifier.issn.pt_BR.fl_str_mv 0100-879X
dc.identifier.doi.none.fl_str_mv 10.1590/1414-431X20143437
identifier_str_mv FREITAS, Bethânia Alves de Avelar et al. Trypan blue exclusion assay by flow cytometry. Braz J Med Biol Res. , v. 47, n. 4, p. 307-15, 2014.
0100-879X
10.1590/1414-431X20143437
url https://www.arca.fiocruz.br/handle/icict/29669
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Associação Brasileira de Divulgação Científica
publisher.none.fl_str_mv Associação Brasileira de Divulgação Científica
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
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