A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green

Autores
Prieto, Daniel; Aparicio, Gonzalo; Morande, Pablo Elías; Zolessi, Flavio
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The increasing need for multiple-labeling of cells and whole organisms for fluorescence microscopy has led to the development of hundreds of fluorophores that either directly recognize target molecules or organelles, or are attached to antibodies or other molecular probes. DNA labeling is essential to study nuclear-chromosomal structure, as well as for gel staining, but also as a usual counterstain in immunofluorescence, FISH or cytometry. However, there are currently few reliable red to far-red-emitting DNA stains that can be used. We describe herein an extremely simple, inexpensive and robust method for DNA labeling of cells and electrophoretic gels using the very well-known histological stain methyl green (MG). MG used in very low concentrations at physiological pH proved to have relatively narrow excitation and emission spectra, with peaks at 633 and 677 nm, respectively, and a very high resistance to photobleaching. It can be used in combination with other common DNA stains or antibodies without any visible interference or bleed-through. In electrophoretic gels, MG also labeled DNA in a similar way to ethidium bromide, but, as expected, it did not label RNA. Moreover, we show here that MG fluorescence can be used as a stain for direct measuring of viability by both microscopy and flow cytometry, with full correlation to ethidium bromide staining. MG is thus a very convenient alternative to currently used red-emitting DNA stains.
Fil: Prieto, Daniel. Universidad de la República; Uruguay. Instituto Pasteur de Montevideo; Uruguay
Fil: Aparicio, Gonzalo. Universidad de la República; Uruguay
Fil: Morande, Pablo Elías. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Pasteur de Montevideo; Uruguay
Fil: Zolessi, Flavio. Universidad de la República; Uruguay. Instituto Pasteur de Montevideo; Uruguay
Materia
Methyl Green
Dna
Flow Cytometry
Confocal Microscopy
Electrophoresis
Fluorescence
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/35891
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/35891
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl greenPrieto, DanielAparicio, GonzaloMorande, Pablo ElíasZolessi, FlavioMethyl GreenDnaFlow CytometryConfocal MicroscopyElectrophoresisFluorescencehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The increasing need for multiple-labeling of cells and whole organisms for fluorescence microscopy has led to the development of hundreds of fluorophores that either directly recognize target molecules or organelles, or are attached to antibodies or other molecular probes. DNA labeling is essential to study nuclear-chromosomal structure, as well as for gel staining, but also as a usual counterstain in immunofluorescence, FISH or cytometry. However, there are currently few reliable red to far-red-emitting DNA stains that can be used. We describe herein an extremely simple, inexpensive and robust method for DNA labeling of cells and electrophoretic gels using the very well-known histological stain methyl green (MG). MG used in very low concentrations at physiological pH proved to have relatively narrow excitation and emission spectra, with peaks at 633 and 677 nm, respectively, and a very high resistance to photobleaching. It can be used in combination with other common DNA stains or antibodies without any visible interference or bleed-through. In electrophoretic gels, MG also labeled DNA in a similar way to ethidium bromide, but, as expected, it did not label RNA. Moreover, we show here that MG fluorescence can be used as a stain for direct measuring of viability by both microscopy and flow cytometry, with full correlation to ethidium bromide staining. MG is thus a very convenient alternative to currently used red-emitting DNA stains.Fil: Prieto, Daniel. Universidad de la República; Uruguay. Instituto Pasteur de Montevideo; UruguayFil: Aparicio, Gonzalo. Universidad de la República; UruguayFil: Morande, Pablo Elías. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Pasteur de Montevideo; UruguayFil: Zolessi, Flavio. Universidad de la República; Uruguay. Instituto Pasteur de Montevideo; UruguaySpringer2014-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/35891Prieto, Daniel; Aparicio, Gonzalo; Morande, Pablo Elías; Zolessi, Flavio; A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green; Springer; Histochemistry And Cell Biology; 142; 3; 9-2014; 335-3450948-6143CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1007/s00418-014-1215-0info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007%2Fs00418-014-1215-0info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:53:04Zoai:ri.conicet.gov.ar:11336/35891instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:53:04.776CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green
title A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green
spellingShingle A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green
Prieto, Daniel
Methyl Green
Dna
Flow Cytometry
Confocal Microscopy
Electrophoresis
Fluorescence
title_short A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green
title_full A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green
title_fullStr A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green
title_full_unstemmed A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green
title_sort A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green
dc.creator.none.fl_str_mv Prieto, Daniel
Aparicio, Gonzalo
Morande, Pablo Elías
Zolessi, Flavio
author Prieto, Daniel
author_facet Prieto, Daniel
Aparicio, Gonzalo
Morande, Pablo Elías
Zolessi, Flavio
author_role author
author2 Aparicio, Gonzalo
Morande, Pablo Elías
Zolessi, Flavio
author2_role author
author
author
dc.subject.none.fl_str_mv Methyl Green
Dna
Flow Cytometry
Confocal Microscopy
Electrophoresis
Fluorescence
topic Methyl Green
Dna
Flow Cytometry
Confocal Microscopy
Electrophoresis
Fluorescence
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The increasing need for multiple-labeling of cells and whole organisms for fluorescence microscopy has led to the development of hundreds of fluorophores that either directly recognize target molecules or organelles, or are attached to antibodies or other molecular probes. DNA labeling is essential to study nuclear-chromosomal structure, as well as for gel staining, but also as a usual counterstain in immunofluorescence, FISH or cytometry. However, there are currently few reliable red to far-red-emitting DNA stains that can be used. We describe herein an extremely simple, inexpensive and robust method for DNA labeling of cells and electrophoretic gels using the very well-known histological stain methyl green (MG). MG used in very low concentrations at physiological pH proved to have relatively narrow excitation and emission spectra, with peaks at 633 and 677 nm, respectively, and a very high resistance to photobleaching. It can be used in combination with other common DNA stains or antibodies without any visible interference or bleed-through. In electrophoretic gels, MG also labeled DNA in a similar way to ethidium bromide, but, as expected, it did not label RNA. Moreover, we show here that MG fluorescence can be used as a stain for direct measuring of viability by both microscopy and flow cytometry, with full correlation to ethidium bromide staining. MG is thus a very convenient alternative to currently used red-emitting DNA stains.
Fil: Prieto, Daniel. Universidad de la República; Uruguay. Instituto Pasteur de Montevideo; Uruguay
Fil: Aparicio, Gonzalo. Universidad de la República; Uruguay
Fil: Morande, Pablo Elías. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Pasteur de Montevideo; Uruguay
Fil: Zolessi, Flavio. Universidad de la República; Uruguay. Instituto Pasteur de Montevideo; Uruguay
description The increasing need for multiple-labeling of cells and whole organisms for fluorescence microscopy has led to the development of hundreds of fluorophores that either directly recognize target molecules or organelles, or are attached to antibodies or other molecular probes. DNA labeling is essential to study nuclear-chromosomal structure, as well as for gel staining, but also as a usual counterstain in immunofluorescence, FISH or cytometry. However, there are currently few reliable red to far-red-emitting DNA stains that can be used. We describe herein an extremely simple, inexpensive and robust method for DNA labeling of cells and electrophoretic gels using the very well-known histological stain methyl green (MG). MG used in very low concentrations at physiological pH proved to have relatively narrow excitation and emission spectra, with peaks at 633 and 677 nm, respectively, and a very high resistance to photobleaching. It can be used in combination with other common DNA stains or antibodies without any visible interference or bleed-through. In electrophoretic gels, MG also labeled DNA in a similar way to ethidium bromide, but, as expected, it did not label RNA. Moreover, we show here that MG fluorescence can be used as a stain for direct measuring of viability by both microscopy and flow cytometry, with full correlation to ethidium bromide staining. MG is thus a very convenient alternative to currently used red-emitting DNA stains.
publishDate 2014
dc.date.none.fl_str_mv 2014-09
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/35891
Prieto, Daniel; Aparicio, Gonzalo; Morande, Pablo Elías; Zolessi, Flavio; A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green; Springer; Histochemistry And Cell Biology; 142; 3; 9-2014; 335-345
0948-6143
CONICET Digital
CONICET
url http://hdl.handle.net/11336/35891
identifier_str_mv Prieto, Daniel; Aparicio, Gonzalo; Morande, Pablo Elías; Zolessi, Flavio; A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green; Springer; Histochemistry And Cell Biology; 142; 3; 9-2014; 335-345
0948-6143
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1007/s00418-014-1215-0
info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007%2Fs00418-014-1215-0
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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