Document Detail


Validation of bromodeoxyuridine immunohistochemistry for localization of S-phase cells in decalcified tissues. A comparative study with tritiated thymidine autoradiography.
MedLine Citation:
PMID:  2228729     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Immunohistochemical detection of the thymidine analogue 5-bromo-2'-deoxyuridine (BrdUrd), which is incorporated by S-phase cells, offers a convenient way of studying the proliferation kinetics of cells in normal skeletal tissues and in bone containing/derived tumours. To assess the validity of using this approach on decalcified, paraffin embedded tissues, the BrdUrd method was compared with tritiated thymidine (3H-TdR) autoradiography, using rat tibiae labelled with both 3H-TdR and BrdUrd, fixed in Carnoy's fluid and decalcified in EDTA, prior to routine paraffin embedding. The distribution of BrdUrd-labelled cells correlated with the sites of cell proliferation in the growing rat tibia. Independent studies with each method on paired serial sections of double-labelled tissue, showed a highly significant correlation (r = 0.81, p less than 0.0003) in the numbers of labelled cells seen in autoradiographs and immunostained sections from the proximal tibial growth plate. Combined BrdUrd immunohistochemistry and 3H-TdR autoradiography showed that the majority of labelled cells in cartilage, bone marrow, and fibrous perichondrium and periosteum had incorporated both labels. These results show that BrdUrd immunohistochemistry is a valid technique for the study of dividing cells in mineralized tissues after decalcification.
Authors:
S S Apte
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Histochemical journal     Volume:  22     ISSN:  0018-2214     ISO Abbreviation:  Histochem. J.     Publication Date:  1990 Aug 
Date Detail:
Created Date:  1990-12-06     Completed Date:  1990-12-06     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0163161     Medline TA:  Histochem J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  401-8     Citation Subset:  IM    
Affiliation:
Nuffield Department of Orthopaedic Surgery, University of Oxford, Nuffield Orthopaedic Centre, Headington.
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MeSH Terms
Descriptor/Qualifier:
Animals
Autoradiography / methods*
Bone Marrow / metabolism
Bone Marrow Cells*
Bromodeoxyuridine / diagnostic use*
Cartilage, Articular / cytology*,  metabolism
Cell Cycle
Cell Division
DNA / metabolism
Growth Plate / cytology*,  metabolism
Immunohistochemistry / methods*
Male
Rats
Rats, Inbred Strains
Thymidine / metabolism
Tibia / cytology*,  metabolism
Tritium / diagnostic use
Chemical
Reg. No./Substance:
10028-17-8/Tritium; 50-89-5/Thymidine; 59-14-3/Bromodeoxyuridine; 9007-49-2/DNA

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


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