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Validation of bromodeoxyuridine immunohistochemistry for localization of S-phase cells in decalcified tissues. A comparative study with tritiated thymidine autoradiography

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Summary

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 both3H-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<0.0003) in the numbers of labelled cells seen in autoradiographs and immunostained sections from the proximal tibial growth plate. Combined BrdUrd immunohistochemistry and3H-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.

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  1. S. S. Apte

    Present address: Department of Anatomy and Cellular Biology, Harvard Medical School, 220 Longwood Avenue, 02115, Boston, MA, USA

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  1. Nuffield Department of Orthopaedic Surgery, University of Oxford, Nuffield Orthopaedic Centre, OX3 7LD, Headington, Oxford, UK

    S. S. Apte

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Apte, S.S. Validation of bromodeoxyuridine immunohistochemistry for localization of S-phase cells in decalcified tissues. A comparative study with tritiated thymidine autoradiography. Histochem J 22, 401–408 (1990). https://doi.org/10.1007/BF01003459

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  • DOI: https://doi.org/10.1007/BF01003459

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