title: Application of the DNA-specific stain methyl green in the fluorescent labeling of embryos.
abstract: 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 fluorophore-conjugated phalloidin.
authors.affiliation: Institut Pasteur de Montevideo.
authors.forename: Daniel
authors.initials: D
authors.lastname: Prieto
authors.affiliation: Institut Pasteur de Montevideo; Sección Biología Celular, Facultad de Ciencias, Universidad de la República.
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Aparicio
authors.affiliation: Institut Pasteur de Montevideo.
authors.forename: Matías
authors.initials: M
authors.lastname: Machado
authors.affiliation: Institut Pasteur de Montevideo; Sección Biología Celular, Facultad de Ciencias, Universidad de la República; fzolessi@fcien.edu.uy.
authors.forename: Flavio R
authors.initials: FR
authors.lastname: Zolessi
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000737:chemistry; D002642:Chick Embryo; D004396:Coloring Agents / Q000737:chemistry*; D004247:DNA / Q000032:analysis*; D004625:Embryo, Nonmammalian / Q000737:chemistry; D004626:Embryology / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D008739:Methyl Green / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods*; D015027:Zebrafish / Q000196:embryology
chemical_list: D004396:Coloring Agents; D005456:Fluorescent Dyes; D008739:Methyl Green; D004247:DNA