Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos
Resumen:
Methyl green has long been known as a histological stain with a specific affinity for DNA, although its fluorescent properties have remained unexplored until recently. In this article, we illustrate the method for preparing a methyl green aqueous stock solution, that when diluted can be used as a very convenient fluorescent nuclear label for fixed cells and tissues. Easy procedures to label whole zebrafish and chick embryos are detailed, and examples of images obtained shown. Methyl green is maximally excited by red light, at 633 nm, and emits with a relatively sharp spectrum that peaks at 677 nm. It is very inexpensive, non-toxic, highly stable in solution and very resistant to photobleaching when bound to DNA. Its red emission allows for unaltered high resolution scanning confocal imaging of nuclei in thick specimens. Finally, this methyl green staining protocol is compatible with other cell staining procedures, such as antibody labeling, or actin filaments labeling with fluorophoreconjugated phalloidin
| 2015 | |
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Developmental biology Issue 99 DNA Methyl green Laser scanning confocal microscopy Fluorescence microscopy Nucleus Wholemount Zebrafish Embryo |
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| Inglés | |
| Universidad de la República | |
| COLIBRI | |
| https://hdl.handle.net/20.500.12008/22052 | |
| Acceso abierto | |
| Licencia Creative Commons Atribución – No Comercial – Sin Derivadas (CC –BY-NC-ND 4.0) |
| _version_ | 1807522780144467968 |
|---|---|
| author | Prieto Mena, Daniel |
| author2 | Aparicio Díaz, Hector Gonzalo Machado, Matías Zolessi, Flavio R. |
| author2_role | author author author |
| author_facet | Prieto Mena, Daniel Aparicio Díaz, Hector Gonzalo Machado, Matías Zolessi, Flavio R. |
| author_role | author |
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| collection | COLIBRI |
| dc.contributor.filiacion.es.fl_str_mv | Prieto Mena, Daniel. IIBCE Aparicio Díaz, Hector Gonzalo. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología Machado, Matías. Instituto Pasteur (Montevideo) Zolessi, Flavio R. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología |
| dc.creator.none.fl_str_mv | Prieto Mena, Daniel Aparicio Díaz, Hector Gonzalo Machado, Matías Zolessi, Flavio R. |
| dc.date.accessioned.none.fl_str_mv | 2019-10-02T22:12:04Z |
| dc.date.available.none.fl_str_mv | 2019-10-02T22:12:04Z |
| dc.date.issued.es.fl_str_mv | 2015 |
| dc.date.submitted.es.fl_str_mv | 20190930 |
| dc.description.abstract.none.fl_txt_mv | Methyl green has long been known as a histological stain with a specific affinity for DNA, although its fluorescent properties have remained unexplored until recently. In this article, we illustrate the method for preparing a methyl green aqueous stock solution, that when diluted can be used as a very convenient fluorescent nuclear label for fixed cells and tissues. Easy procedures to label whole zebrafish and chick embryos are detailed, and examples of images obtained shown. Methyl green is maximally excited by red light, at 633 nm, and emits with a relatively sharp spectrum that peaks at 677 nm. It is very inexpensive, non-toxic, highly stable in solution and very resistant to photobleaching when bound to DNA. Its red emission allows for unaltered high resolution scanning confocal imaging of nuclei in thick specimens. Finally, this methyl green staining protocol is compatible with other cell staining procedures, such as antibody labeling, or actin filaments labeling with fluorophoreconjugated phalloidin |
| dc.format.mimetype.es.fl_str_mv | application/pdf |
| dc.identifier.citation.es.fl_str_mv | Prieto, D., et al. Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos. Journal of Visualized Experiments. 2015, (99), e52769. doi:10.3791/52769 |
| dc.identifier.doi.es.fl_str_mv | 10.3791/52769 |
| dc.identifier.issn.es.fl_str_mv | 1940-087X |
| dc.identifier.uri.none.fl_str_mv | https://hdl.handle.net/20.500.12008/22052 |
| dc.language.iso.none.fl_str_mv | en eng |
| dc.publisher.es.fl_str_mv | JOVE |
| dc.relation.ispartof.es.fl_str_mv | Journal of Visualized Experiments. 2015, (99), e52769 |
| dc.rights.license.none.fl_str_mv | Licencia Creative Commons Atribución – No Comercial – Sin Derivadas (CC –BY-NC-ND 4.0) |
| dc.rights.none.fl_str_mv | info:eu-repo/semantics/openAccess |
| dc.source.none.fl_str_mv | reponame:COLIBRI instname:Universidad de la República instacron:Universidad de la República |
| dc.subject.es.fl_str_mv | Developmental biology Issue 99 DNA Methyl green Laser scanning confocal microscopy Fluorescence microscopy Nucleus Wholemount Zebrafish Embryo |
| dc.title.none.fl_str_mv | Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos |
| dc.type.es.fl_str_mv | Artículo |
| dc.type.none.fl_str_mv | info:eu-repo/semantics/article |
| dc.type.version.none.fl_str_mv | info:eu-repo/semantics/publishedVersion |
| description | Methyl green has long been known as a histological stain with a specific affinity for DNA, although its fluorescent properties have remained unexplored until recently. In this article, we illustrate the method for preparing a methyl green aqueous stock solution, that when diluted can be used as a very convenient fluorescent nuclear label for fixed cells and tissues. Easy procedures to label whole zebrafish and chick embryos are detailed, and examples of images obtained shown. Methyl green is maximally excited by red light, at 633 nm, and emits with a relatively sharp spectrum that peaks at 677 nm. It is very inexpensive, non-toxic, highly stable in solution and very resistant to photobleaching when bound to DNA. Its red emission allows for unaltered high resolution scanning confocal imaging of nuclei in thick specimens. Finally, this methyl green staining protocol is compatible with other cell staining procedures, such as antibody labeling, or actin filaments labeling with fluorophoreconjugated phalloidin |
| eu_rights_str_mv | openAccess |
| format | article |
| id | COLIBRI_a3f0c1716bd2dd186065da857680c69c |
| identifier_str_mv | Prieto, D., et al. Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos. Journal of Visualized Experiments. 2015, (99), e52769. doi:10.3791/52769 1940-087X 10.3791/52769 |
| instacron_str | Universidad de la República |
| institution | Universidad de la República |
| instname_str | Universidad de la República |
| language | eng |
| language_invalid_str_mv | en |
| network_acronym_str | COLIBRI |
| network_name_str | COLIBRI |
| oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/22052 |
| publishDate | 2015 |
| reponame_str | COLIBRI |
| repository.mail.fl_str_mv | mabel.seroubian@seciu.edu.uy |
| repository.name.fl_str_mv | COLIBRI - Universidad de la República |
| repository_id_str | 4771 |
| rights_invalid_str_mv | Licencia Creative Commons Atribución – No Comercial – Sin Derivadas (CC –BY-NC-ND 4.0) |
| spelling | Prieto Mena, Daniel. IIBCEAparicio Díaz, Hector Gonzalo. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de BiologíaMachado, Matías. Instituto Pasteur (Montevideo)Zolessi, Flavio R. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología2019-10-02T22:12:04Z2019-10-02T22:12:04Z201520190930Prieto, D., et al. Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos. Journal of Visualized Experiments. 2015, (99), e52769. doi:10.3791/527691940-087Xhttps://hdl.handle.net/20.500.12008/2205210.3791/52769Methyl green has long been known as a histological stain with a specific affinity for DNA, although its fluorescent properties have remained unexplored until recently. In this article, we illustrate the method for preparing a methyl green aqueous stock solution, that when diluted can be used as a very convenient fluorescent nuclear label for fixed cells and tissues. Easy procedures to label whole zebrafish and chick embryos are detailed, and examples of images obtained shown. Methyl green is maximally excited by red light, at 633 nm, and emits with a relatively sharp spectrum that peaks at 677 nm. It is very inexpensive, non-toxic, highly stable in solution and very resistant to photobleaching when bound to DNA. Its red emission allows for unaltered high resolution scanning confocal imaging of nuclei in thick specimens. Finally, this methyl green staining protocol is compatible with other cell staining procedures, such as antibody labeling, or actin filaments labeling with fluorophoreconjugated phalloidinMade available in DSpace on 2019-10-02T22:12:04Z (GMT). No. of bitstreams: 5 10379152769.pdf: 946080 bytes, checksum: a2ac6d0023cc0c375bf02ce79804398e (MD5) license_text: 38300 bytes, checksum: 098d76773c7b7afafb04cabc04ea8a56 (MD5) license_url: 47 bytes, checksum: 966d4a1cc97b2c4389b5142dd97d3c7f (MD5) license_rdf: 9754 bytes, checksum: ffcba5f515f45166c8d3bb6aa02e3123 (MD5) license.txt: 4194 bytes, checksum: 7f2e2c17ef6585de66da58d1bfa8b5e1 (MD5) Previous issue date: 2015application/pdfenengJOVEJournal of Visualized Experiments. 2015, (99), e52769Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad De La República. (Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución – No Comercial – Sin Derivadas (CC –BY-NC-ND 4.0)Developmental biologyIssue 99DNAMethyl greenLaser scanning confocal microscopyFluorescence microscopyNucleusWholemountZebrafishEmbryoApplication of the DNA-Specific stain methyl green in the fluorescent labeling of embryosArtículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionreponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaPrieto Mena, DanielAparicio Díaz, Hector GonzaloMachado, MatíasZolessi, Flavio 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- Universidad de la Repúblicafalse |
| spellingShingle | Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos Prieto Mena, Daniel Developmental biology Issue 99 DNA Methyl green Laser scanning confocal microscopy Fluorescence microscopy Nucleus Wholemount Zebrafish Embryo |
| status_str | publishedVersion |
| title | Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos |
| title_full | Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos |
| title_fullStr | Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos |
| title_full_unstemmed | Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos |
| title_short | Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos |
| title_sort | Application of the DNA-Specific stain methyl green in the fluorescent labeling of embryos |
| topic | Developmental biology Issue 99 DNA Methyl green Laser scanning confocal microscopy Fluorescence microscopy Nucleus Wholemount Zebrafish Embryo |
| url | https://hdl.handle.net/20.500.12008/22052 |
