skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Enhanced detection of fluorescence quenching in labeled cells

Abstract

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.

Inventors:
;
Publication Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
OSTI Identifier:
6293007
Patent Number(s):
PATENTS-US-A7126155
Application Number:
ON: DE89010955
Assignee:
Dept. of Energy
DOE Contract Number:  
W-7405-ENG-36
Resource Type:
Patent
Resource Relation:
Other Information: Paper copy only, copy does not permit microfiche production
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 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

Citation Formats

Crissman, H. A., and Steinkamp, J. A. Enhanced detection of fluorescence quenching in labeled cells. United States: N. p., 1987. Web.
Crissman, H. A., & Steinkamp, J. A. Enhanced detection of fluorescence quenching in labeled cells. United States.
Crissman, H. A., and Steinkamp, J. A. Mon . "Enhanced detection of fluorescence quenching in labeled cells". United States. https://www.osti.gov/servlets/purl/6293007.
@article{osti_6293007,
title = {Enhanced detection of fluorescence quenching in labeled cells},
author = {Crissman, H. A. and Steinkamp, J. A.},
abstractNote = {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.},
doi = {},
url = {https://www.osti.gov/biblio/6293007}, journal = {},
number = ,
volume = ,
place = {United States},
year = {1987},
month = {11}
}