• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. Sep 1983; 80(18): 5573–5577.
PMCID: PMC384300

Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine.

Abstract

We have developed a procedure for simultaneous flow cytometric measurement of cellular DNA content and amount of BrdUrd incorporated into cellular DNA. Propidium iodide was used as a fluorescent probe for total cellular DNA and a monoclonal antibody against BrdUrd was used as a probe for BrdUrd incorporated into DNA. Fluorescein-labeled goat anti-mouse antibody was used to fluorescently label the bound anti-BrdUrd probe. Bivariate DNA/BrdUrd distributions measured for Chinese hamster ovary cells labeled for 30 min with BrdUrd clearly show the G1-and G2M-phase cells to have low BrdUrd-linked fluorescence and the S-phase cells to have high BrdUrd-linked fluorescence. Cell cycle traverse rates were estimated for Chinese hamster ovary cells from bivariate distributions measured for samples taken periodically after pulse labeling with BrdUrd. Bivariate DNA/BrdUrd distributions were also applied in the analysis of the response of C3H murine bone marrow cells to treatment in vivo with 1-beta-D-arabinofuranosylcytosine (araC). Bivariate distributions were measured for bone marrow cells taken from mice that were pulse labeled with BrdUrd at various times after treatment with araC. The resulting DNA/BrdUrd sequences show the kinetics of recovery from araC and allow discrimination of the araC sterilized cells.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Hart JS, George SL, Frei E, 3rd, Bodey GP, Nickerson RC, Freireich EJ. Prognotic significance of pretreatment proliferative activity in adult acute leukemia. Cancer. 1977 Apr;39(4):1603–1617. [PubMed]
  • Barranco SC, May JT, Boerwinkle W, Nichols S, Hokanson KM, Schumann J, Göhde W, Bryant J, Guseman LF. Enhanced cell killing through the use of cell kinetics-directed treatment schedules for two-drug combinations in vitro. Cancer Res. 1982 Jul;42(7):2894–2898. [PubMed]
  • QUASTLER H, SHERMAN FG. Cell population kinetics in the intestinal epithelium of the mouse. Exp Cell Res. 1959 Jun;17(3):420–438. [PubMed]
  • Pallavicini MG, Gray JW, Folstad LJ. Quantitative analysis of the cytokinetic response of KHT tumors in vivo to 1-beta-D-arabinofuranosylcytosine. Cancer Res. 1982 Aug;42(8):3125–3131. [PubMed]
  • Painter RB, Young BR. Formation of nascent DNA molecules during inhibition of replicon initiation in mammalian cells. Biochim Biophys Acta. 1976 Jan 19;418(2):146–153. [PubMed]
  • Simpson-Herren L, Sanford AH, Holmquist JP, Springer TA, Lloyd HH. Ambiguity of the thymidine index. Cancer Res. 1976 Dec;36(12):4705–4709. [PubMed]
  • Gratzner HG. Monoclonal antibody to 5-bromo- and 5-iododeoxyuridine: A new reagent for detection of DNA replication. Science. 1982 Oct 29;218(4571):474–475. [PubMed]
  • Gray JW, Pallavicini MG, George YS, Groppi V, Look M, Dean PN. Rates of incorporation of radioactive molecules during the cell cycle. J Cell Physiol. 1981 Aug;108(2):135–144. [PubMed]
  • Gratzner HG, Leif RC. An immunofluorescence method for monitoring DNA synthesis by flow cytometry. Cytometry. 1981 May;1(6):385–393. [PubMed]
  • Latt SA, George YS, Gray JW. Flow cytometric analysis of bromodeoxyuridine-substituted cells stained with 33258 Hoechst. J Histochem Cytochem. 1977 Jul;25(7):927–934. [PubMed]
  • Böhmer RM, Ellwart J. Cell cycle analysis by combining the 5-bromodeoxyuridine/33258 Hoechst technique with DNA-specific ethidium bromide staining. Cytometry. 1981 Jul;2(1):31–34. [PubMed]
  • Gray JW, Coffino P. Cell cycle analysis by flow cytometry. Methods Enzymol. 1979;58:233–248. [PubMed]
  • Traganos F, Darzynkiewicz Z, Sharpless T, Melamed MR. Cytofluorometric studies on conformation of nucleic acids in situ. I. Restriction of acridine orange binding by chromatin proteins. J Histochem Cytochem. 1976 Jan;24(1):40–48. [PubMed]
  • LePecq JB, Paoletti C. A fluorescent complex between ethidium bromide and nucleic acids. Physical-chemical characterization. J Mol Biol. 1967 Jul 14;27(1):87–106. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...