Double labeling with iodo- and bromodeoxyuridine for cell kinetic studies

S. Shibui, T. Hoshino, M. Vanderlaan, J. W. Gray

Research output: Contribution to journalArticlepeer-review

79 Scopus citations


The rate of progression through the cell cycle was determined in five human glioma cell lines by a new sequential immunohistochemical staining technique. The cells were labeled first with iododeoxyuridine (IdUrd) for 1-3 hr and then with bromodeoxyuridine (BrdUrd) for 30 min. Labeled cells were identified with Br-3, a monoclonal antibody that recognizes only BrdUrd, and with IU-4, an antibody that recognizes both IdUrd and BrdUrd. Each slide was stained sequentially, first with the immunoperoxidase method for Br-3 and then with the alkaline phosphatase-anti-alkaline phosphatase method for IU-4. Cells that were positive only for IU-4 represented the fraction of S-phase cells that passed into the G2 phase during the period of incubation with IdUrd. The rates of progression measured by this method were constant in each cell line and resulted in smaller standard errors than were obtained by measurements from specimens stained singly for IdUrd and BrdUrd in different slides. The duration of the S-phase calculated from this fraction in the five cell lines ranged from 8-13 hr; the estimated potential doubling times were 25-32 hr and were very similar to the actual doubling times.

Original languageEnglish (US)
Pages (from-to)1007-1011
Number of pages5
JournalJournal of Histochemistry and Cytochemistry
Issue number7
StatePublished - 1989
Externally publishedYes

ASJC Scopus subject areas

  • Anatomy
  • Histology


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