Document Detail


Measurement of DNA content and of tritiated thymidine and bromodeoxyuridine incorporation by the same cells.
MedLine Citation:
PMID:  8354883     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We tested a method of measuring DNA content (Feulgen) and tritiated thymidine ([3H]-T) and bromodeoxyuridine (BrdU) incorporation by the same cell. Initial experiments showed that Feulgen hydrolysis denatured the DNA of fixed cells sufficiently to allow detection of incorporated BrdU with monoclonal antibodies. MCa-11 cells were then double-labeled with [3H]-T and BrdU, placed on slides, and Feulgen stained. Next, absorption cytometry was performed to measure the DNA content of randomly selected cells. Feulgen staining and the development and removal of either the [3H]-T or the BrdU grains after DNA measurements did not interfere with subsequent detection of the grains from the other label, and BrdU and [3H]-T can be used reliably in combination for identification of S-phase cells. This method may eventually allow the use of microscope-based image analysis to selectively measure the DNA contents and the BrdU/[3H]-T labeling of non-transformed stromal and cancer cells in solid tumors, thereby providing new insights into the growth kinetics of these heterogeneous cell populations.
Authors:
P Lin; D C Allison
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society     Volume:  41     ISSN:  0022-1554     ISO Abbreviation:  J. Histochem. Cytochem.     Publication Date:  1993 Sep 
Date Detail:
Created Date:  1993-09-21     Completed Date:  1993-09-21     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  9815334     Medline TA:  J Histochem Cytochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1435-9     Citation Subset:  IM    
Affiliation:
Department of Surgery, Johns Hopkins Medical Institutes, Baltimore, Maryland.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies, Monoclonal
Autoradiography
Bromodeoxyuridine / metabolism*
Cell Division
DNA, Neoplasm / analysis*,  metabolism
Female
Image Processing, Computer-Assisted
Mammary Neoplasms, Experimental* / chemistry,  metabolism,  pathology
Mice
S Phase
Thymidine / metabolism*
Tritium
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/DNA, Neoplasm; 10028-17-8/Tritium; 50-89-5/Thymidine; 59-14-3/Bromodeoxyuridine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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