Rapid staining methods for analysis of deocyribonucleic acid and protein in mammalian cells

Journal Article · Thu Jan 01 00:00:00 EST 1976 · J. Histochem. Cytochem.; (United States)

DOI:https://doi.org/10.1177/24.1.56392· OSTI ID:7256597

Crissman, H A; Oka, M S; Steinkamp, J A

Quantitative fluorescent staining and analysis of cellular deoxyribonucleic acid (DNA) were accomplished using three groups of reagents having different mechanisms of action for DNA binding. These reagents included the fluorescent antitumor antibiotics mithramycin, chromomycin A/sub 3/ and olivomycin; the Feulgen reagents acriflavine and flavophosphine N; and the intercalating dyes ethidium bromide and propidium iodide. Propidium iodide (PI) was used in combination with fluorescein isothiocyanate (FITC) to stain both cellular DNA and protein, respectively. Multiparameter analysis of PI/FITC-stained cells provided a direct correlation of DNA and protein for cells in all stages of the cell cycle. Nuclear-to-cytoplasmic ratio determinations were also performed on PI/FITC-stained cells by analysis of the time duration of the red (DNA) and green (protein) fluorescence signals from each cell. These staining and analysis techniques provide alternative methods for directly determining the quantitative relationship between cellular DNA and protein and will be extremely useful in investigations where fluctuations of these parameters are of importance for assessing experimental results.

• Research Organization: Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
• OSTI ID: 7256597
• Journal Information: J. Histochem. Cytochem.; (United States), Vol. 24:1
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
CYTOCHEMISTRY
DNA
CHEMICAL ANALYSIS
PROTEINS
ACRIFLAVINE
ANTIBIOTICS
CELL NUCLEI
CYTOPLASM
FEULGEN METHOD
FLUORESCEIN
FLUORESCENCE
STAINS
ACRIDINES
AMINES
AROMATICS
AZINES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
DRUGS
DYES
FLAVINES
HETEROCYCLIC COMPOUNDS
HYDROXY ACIDS
HYDROXY COMPOUNDS
LUMINESCENCE
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHENOLS
POLYPHENOLS
PYRIDINES
550300* - Cytology
550200 - Biochemistry