PUBLICATION
Application of the DNA-Specific Stain Methyl Green in the Fluorescent Labeling of Embryos
- Authors
- Prieto, D., Aparicio, G., Machado, M., Zolessi, F.R.
- ID
- ZDB-PUB-150521-2
- Date
- 2015
- Source
- Journal of visualized experiments : JoVE (99): e52769 (Journal)
- Registered Authors
- Aparicio, Gonzalo, Prieto, Daniel, Zolessi, Flavio
- Keywords
- none
- MeSH Terms
-
- Animals
- Cell Nucleus/chemistry
- Chick Embryo
- Coloring Agents/chemistry*
- DNA/analysis*
- Embryo, Nonmammalian/chemistry
- Embryology/methods*
- Fluorescent Dyes/chemistry*
- Methyl Green/chemistry*
- Staining and Labeling/methods*
- Zebrafish/embryology
- PubMed
- 25993383 Full text @ J. Vis. Exp.
Citation
Prieto, D., Aparicio, G., Machado, M., Zolessi, F.R. (2015) Application of the DNA-Specific Stain Methyl Green in the Fluorescent Labeling of Embryos. Journal of visualized experiments : JoVE. (99):e52769.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping