Flow microfluorometric and spectrophotofluorometric analysis of DNA staining in mammalian cells
The effects of pH, ionic strength, stain concentration, and magnesium concentration on DNA staining with the antibiotics mithramycin, chromomycin A3, and olivomycin were examined with DNA in solution and in mammalian cells. Ethanol-fixed Chinese hamster cells (line CHO) stained with mithramycin solution in the pH range 4.0 to 10.0 and analyzed by flow microfluorometry (FMF) showed only a slight increase in fluorescence intensity from pH 4.0 to 8.0. Above this range, there was a more dramatic increase in intensity of stained cells, and at pH 10.0 the flouorescence intensity was 1-1/2 times greater than cells stained at pH 4.5. The resolution in DNA distribution patterns also improved as a function of increasing pH, corresponding with a marked decrease in the coefficient of variation (CV). However, the distribution of cells in various phases of the cell cycle remained essentially the same over the pH ranges tested.
- Research Organization:
- Los Alamos Scientific Lab., NM (USA)
- DOE Contract Number:
- W-7405-ENG-36
- OSTI ID:
- 7098140
- Report Number(s):
- LA-UR-77-365; CONF-770324-3
- Country of Publication:
- United States
- Language:
- English
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