7‐amino‐actinomycin D as a specific fluorophore for DNA content analysis by laser flow cytometry

A technique for DNA amount determination by flow cytometry based on the use of 7‐amino‐actinomycin D (7‐amino‐AMD), a fluorescent analogue of antibiotic actinomycin has been investigated, and a particular staining procedure has been developed. The procedure includes short fixation in 70% ethanol and staining for 20 min in 10‐5M solution of 7‐amino‐AMD at pH7. The results of DNA content measurements are very reproducible. The histograms obtained have a coefficient of variation less than 3%. The absorption maximum of the complex of 7‐amino‐AMD with DNA is situated in the green spectrum region, making this stain particularly suitable for argon laser flow cytometry.