Bromodeoxyuridine: a diagnostic tool in biology and medicine, Part I: Historical perspectives, histochemical methods and cell kinetics

Abstract

Bromodeoxyuridine (BrdUrd), a thymidine analogue incorporated into DNA, can be quantified by fluorescent or chromophoric quenching of dyes bound to DNA or with antibodies to BrdUrd. These technologies have been used since the 1970s as tools for measuring DNA synthesis in isolated chromosomes and in cells and tissues. This paper is Part I of a three-part comprehensive review of the literature over the last 20 years (to the end of 1993) describing the histochemical methods for measuring BrdUrd in cells and tissues. Fixation, denaturation and staining procedures are compared for quantifying BrdUrd for microscopy and flow cytometry. Non-immunochemical methods related to the quenching of fluorescent DNA stains by BrdUrd are also described. Methods are described for the comparative assay of cell kinetic parameters by tritiated thymidine and bromodeoxyuridine. The multivariate BrdUrd/DNA assay of Ts and Tc, and a comparison of recent methods based on the single biopsy bivariate analysis of Tpot, is presented. Recent developments in the use of double halopyrimidine label to determine kinetic parameters are also reviewed.

Spatiotemporal patterns of secretomotor neuron generation in the parvicellular neuroendocrine system 1Published on the World Wide Web on 3 June 1997. 1

“…The advantages of birthdating with BrdU, as compared to [ 3 H]thymidine, are very low background, intranuclear labeling [21,22,41], and visualization the uridine analog with immunofluorescence, which allows easy, simultaneous immunofluorescence localization of a neurotransmitter and detection of fluorescent axonal tracers. The major limitation of BrdU birthdating is that the immunohistochemical label is not as readily quantifiable as the unitary silver grains associated with [ 3 H]thymidine.…”

“…In the present study, birthdates of the 6 best characterized PCNs have been examined with the bromodeoxyuridine (BrdU) method [21,22,34,41]. Nuclear incorporation of BrdU was detected with immunofluorescence, allowing simultaneous immunofluorescence detection of characteristic neurotransmitters.…”

The antiproliferative activity of all-trans-retinoic acid catabolites and isomers is differentially modulated by liarozole-fumarate in MCF-7 human breast cancer cells

“…Cell proliferation was measured using BrdU incorporation, which is considered an accurate method because it is a direct assay of DNA synthesis (Dolbeare, 1995). After 7 days of culture with retinoids as described above, MCF-7 cells were labelled with 100 gM BrdU for 2.5 h and fixed.…”

All-trans-retinoic acid metabolites significantly inhibit the proliferation of MCF-7 human breast cancer cells in vitro

“…To clarify these points, we studied the antiproliferative activity of ATRA, its physiological catabolites 4-oxo-ATRA and 5,6-epoxy-ATRA and isomers 9-cis-RA and 13-cis-RA in MCF-7 cells using a bromodeoxyuridine incorporation assay. Bromodeoxyuridine labelling is considered an accurate method to measure cell proliferation because it is a direct assay of DNA synthesis (Dolbeare, 1995). Furthermore, the antiproliferative activities could only be properly studied in a steroid-and retinoid-free medium, because of the presence of endogenous retinoids in serum.…”