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Title: Enhanced detection of fluorescence quenching in labeled cells

Abstract

A method is provided for quantifying BrdU labeled DNA in cells. The BrdU is incorporated into 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 that 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 substracted 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.

Inventors:
 [1];  [1]
  1. Los Alamos, NM
Issue Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
OSTI Identifier:
868152
Patent Number(s):
5084378
Assignee:
United States of America as represented by United States (Washington, DC)
Patent Classifications (CPCs):
Y - NEW / CROSS SECTIONAL TECHNOLOGIES Y10 - TECHNICAL SUBJECTS COVERED BY FORMER USPC Y10T - TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
DOE Contract Number:  
W-7405-ENG-36
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
enhanced; detection; fluorescence; quenching; labeled; cells; method; provided; quantifying; brdu; dna; incorporated; stained; fluorochrome; quenchable; preferably; thymidine; base; halogen; analogue; hoechst; substantially; uneffected; selected; consisting; mithramycin; chromomycin; a3; olivomycin; propidium; iodide; ethidium; bromine; fluorochromes; measured; obtain; output; signals; respectively; signal; substracted; difference; functionally; related; quantity; technique; particularly; useful; synthesis; s-phase; cell; cycle; difference signal; labeled dna; functionally related; output signals; particularly useful; output signal; cell cycle; propidium iodide; /435/436/

Citation Formats

Crissman, Harry A, and Steinkamp, John A. Enhanced detection of fluorescence quenching in labeled cells. United States: N. p., 1992. Web.
Crissman, Harry A, & Steinkamp, John A. Enhanced detection of fluorescence quenching in labeled cells. United States.
Crissman, Harry A, and Steinkamp, John A. Wed . "Enhanced detection of fluorescence quenching in labeled cells". United States. https://www.osti.gov/servlets/purl/868152.
@article{osti_868152,
title = {Enhanced detection of fluorescence quenching in labeled cells},
author = {Crissman, Harry A and Steinkamp, John A},
abstractNote = {A method is provided for quantifying BrdU labeled DNA in cells. The BrdU is incorporated into 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 that 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 substracted 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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1992},
month = {1}
}

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