Flow cytometric analysis of bromodeoxyuridine-substituted cells stained with 33258 Hoechst

Journal Article · Fri Dec 31 23:00:00 EST 1976 · J. Histochem. Cytochem.; (United States)

DOI:https://doi.org/10.1177/25.7.70460· OSTI ID:7192202

Latt, S A ^[1]; George, Y S; Gray, J W

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1. Childrens Hospital Medical Center, Boston, MA

The paper describes a flow-cytometric application of the quenching of fluorescence from 33258 Hoechst stained Chinese hamster ovary-line cells due to the incorporation of 5-bromo-deoxyuridine (BrdU) into the cellular deoxyribonucleic acid. Cells were grown for 24 hr in medium containing BrdU in concentrations ranging from 1 x 10/sup -8/ to 1 x 10/sup -4/ M. For each concentration the average fluorescence was measured as determined by flow cytometry, the extent of BrdU substitution and the effect of the BrdU on cell growth. BrdU concentrations of 1 x 10/sup -5/ M resulted in sufficient substitution to quench the fluorescence from 33258 Hoechst by a factor of 4, allowing discrimination between cycling and noncycling cells. The extent of BrdU substitution after growth for 24 hr in this concentration of BrdU was 64%. These data indicate the feasibility of detecting deoxyribonucleic acid synthesis in whole cells using the 33258 Hoechst-BrdU methodology.

• DOE Contract Number: W-7405-ENG-48
• OSTI ID: 7192202
• Journal Information: J. Histochem. Cytochem.; (United States), Journal Name: J. Histochem. Cytochem.; (United States) Vol. 25:7; ISSN JHCYA
• Country of Publication: United States
• Language: English

Related Subjects

550300* -- Cytology
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
ANTIMETABOLITES
AZINES
BIOSYNTHESIS
BODY
BROMOURACILS
BUDR
CELL FLOW SYSTEMS
COMPLEXES
CYTOLOGICAL TECHNIQUES
DNA
DRUGS
DYES
FEMALE GENITALS
FLUORESCENCE
GONADS
HAMSTERS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
LUMINESCENCE
MAMMALS
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC BROMINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
OVARIES
PYRIMIDINES
QUENCHING
RIBOSIDES
RODENTS
SYNTHESIS
URACILS
VERTEBRATES