Histochemistry

, Volume 90, Issue 1, pp 31–35 | Cite as

Detection of cell proliferation in pig testis and intestine sections using monoclonal anti-bromodeoxyuridine antibody and immunogold silver staining

  • D. Dombrowicz
  • P. Delahaut
  • A. Danguy
  • J. Closset
  • G. Hennen
Article

Summary

For the first time a monoclonal antibody against 5-bromodeoxyuridine was used to detect cell proliferation in pig testis and intestine sections. The influence of several parameters such as mode of injection, addition of thymidine biosynthesis inhibitor, tissue fixation, hydrolysis and revelation was examined. The technique of choice consisted in intravenously injecting the animals with 50 mg/kg BUdR added to 10 mg/kg FUdR 2 h before tissue collection and Bouin fixation; hydrolysis of sections was performed by HCl 4N: Ethanol 70° (1:1 v/v); revelation of BUdR was made by a secondary antibody linked to colloidal gold particles, followed by a silver enhancement step. The data were superior when compared to those obtained by direct immunofluorescence and by the PAP technique. The described method is convenient and sensitive, provides an intense nuclear labelling without background and allows simultaneous examination of histology. The advantages over the technique using tritiated thymidine are particularly obvious when fast screening of numerous samples is required or when new experimental protocols are developping.

Keywords

Thymidine Tritiated Thymidine Biosynthesis Inhibitor Colloidal Gold Particle Fast Screening 

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Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • D. Dombrowicz
    • 1
  • P. Delahaut
    • 2
  • A. Danguy
    • 3
  • J. Closset
    • 1
  • G. Hennen
    • 1
  1. 1.Endocrinologie Expérimentale et Clinique, Faculté de MédecineUniversité de LiègeSart-Tilman LiègeBelgique
  2. 2.Centre Provincial d'Economie RuraleMarloieBelgique
  3. 3.Histologie, Faculté de MédecineUniversité Libre de BruxellesBruxellesBelgique

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