Determination of the fraction of S-phase cells in root meristems using bromodeoxyuridine labeling

José P. Hervás, J. de la Flor, M. C. Santa-Cruz

Research output: Contribution to journalArticleResearchpeer-review

3 Citations (Scopus)


The use of bromodeoxyuridine (BrdU) and subsequent immunocytochemical visualization for studying cell proliferation in plant meristems was investigated in Allium cepa L. root-tips. We describe the optimization of an indirect immunoperoxidase method for detecting incorporation of this DNA precursor in pulse-labeled cells. The basic object of this study is to quantify the extent to which the fraction of S-phase cells can reliably be estimated in asynchronous populations. A matrix of parallel labeling schedules with tritiated-thymidine or BrdU was developed, and the labeling indices provided by autoradiography or immunocytochemistry were compared. Thus, 0.5 mM BrdU assured saturation S-phase labeling after an exposure time of 30 min, and the mean length of the S-phase determined under such conditions was similar to that previously reported for this plant system. Interestingly, Feulgen staining did not interfere with subsequent detection of the BrdU probe. This allowed comparative evaluations of the nuclear DNA content by Feulgen-microdensitometry and the position of a given cell in G1, S or G2 compartments. We also explored the possibility of quantifying BrdU-incorporation in single nuclei by densitometry measurement of the peroxidase label.
Original languageEnglish
Pages (from-to)145-152
JournalBiotechnic and Histochemistry
Issue number3
Publication statusPublished - 1 Jan 2002


  • Allium cepa
  • Bromodeoxyuridine
  • Cell cycle
  • Immunoperoxidase
  • Meristems
  • Microdensitometry
  • S-phase


Dive into the research topics of 'Determination of the fraction of S-phase cells in root meristems using bromodeoxyuridine labeling'. Together they form a unique fingerprint.

Cite this