Enhanced detection of fluorescence quenching in labeled cells
A method is provided for quantifying BrdU labeled DNA in cells. The BrdU is substituted onto the DNA and the DNA is stained with a first fluorochrome having a fluorescence which is quenchable by BrdU. The first fluorochrome is preferably a thymidine base halogen analogue, such as a Hoechst fluorochrome. The DNA is then stained with a second fluorochrome having a fluorescence which is substantially uneffected by BrdU. The second fluorochrome may be selected from the group consisting of mithramycin, chromomycin A3, olivomycin, propidium iodide and ethidium bromine. The fluorescence from the first and second fluorochromes is then measured to obtain first and second output signals, respectively. The first output signal is subtracted from the second output signal to obtain a difference signal which is functionally related to the quantity of BrdU incorporated into DNA. The technique is particularly useful for quantifying the synthesis of DNA during the S-phase of the cell cycle. 2 figs.
- Research Organization:
- Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
- DOE Contract Number:
- W-7405-ENG-36
- Assignee:
- Dept. of Energy
- Patent Number(s):
- PATENTS-US-A7126155
- Application Number:
- ON: DE89010955
- OSTI ID:
- 6293007
- Resource Relation:
- Other Information: Paper copy only, copy does not permit microfiche production
- Country of Publication:
- United States
- Language:
- English
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FLUORESCENCE SPECTROSCOPY
EVALUATION
STAINS
LABELLING
BUDR
DNA
DNA SEQUENCING
INVENTIONS
QUANTITATIVE CHEMICAL ANALYSIS
ANTIMETABOLITES
AZINES
BROMOURACILS
CHEMICAL ANALYSIS
DRUGS
EMISSION SPECTROSCOPY
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC BROMINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PYRIMIDINES
RIBOSIDES
SPECTROSCOPY
STRUCTURAL CHEMICAL ANALYSIS
URACILS
550200* - Biochemistry