PUBLICATION

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

Authors
Prieto, D., Aparicio, G., Morande, P.E., Zolessi, F.R.
ID
ZDB-PUB-151110-12
Date
2014
Source
Histochemistry and cell biology   142: 335-345 (Journal)
Registered Authors
Aparicio, Gonzalo, Prieto, Daniel, Zolessi, Flavio
Keywords
Methyl green, DNA, Flow cytometry, Confocal microscopy, Electrophoresis, Fluorescence
MeSH Terms
  • Animals
  • DNA/analysis*
  • DNA/chemistry
  • Fluorescent Dyes/chemistry*
  • Methyl Green/chemistry*
  • Microscopy, Fluorescence
  • Staining and Labeling/economics*
  • Staining and Labeling/methods*
  • Time Factors
  • Zebrafish/embryology
PubMed
24671497 Full text @ Histochem. Cell Biol.
Abstract
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.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping