Advertisement

Cell Cycle Analysis of Asynchronous Populations

Protocol
  • 1.2k Downloads
Part of the Methods in Molecular Biology™ book series (MIMB, volume 91)

Abstract

Measurement of a DNA histogram can be achieved by fixing or permeabilizing cells and staining them with a DNA-binding dye, such as propidium iodide (PI). The histogram will yield the percentage of cells in the G1, S, and G2/M phases of the cell cycle (1). Although some inferences about the movement of cells through the cycle may be drawn, the information gained is essentially static. For example, it is not known whether a cell with S-phase DNA content is actually synthesizing DNA; also the presence of subpopulations with different cycle times cannot be detected.

Keywords

Propidium Iodide Propidium Iodide Blue Fluorescence Staining Buffer Medulloblastoma Cell Line 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in to check access.

References

  1. 1.
    Ormerod, M G (1994) Analysis of DNA: general methods, in Flow Cytometry A Practical Approach, 2nd ed. (Ormerod, M. G., ed), IRL Press, Oxford, UK, pp. 118–135.Google Scholar
  2. 2.
    Dolbeare, F. A., Gratzner, H. G, Pallavicini, M. G., and Gray, J. W. (1983) Flow cytometric measurement of total DNA content and incorporated bromodeoxyundine. Proc Natl Acad. Sci. USA 80, 5573–5577.PubMedCrossRefGoogle Scholar
  3. 3.
    Begg, A. C, McNally, N. J., Shrieve, D. C, and Karcher, H. (1985) A method to measure the duration of DNA synthesis and potential doubling time from a single sample. Cytometry 6, 620–626.PubMedCrossRefGoogle Scholar
  4. 4.
    Wilson, G. D. (1994) Analysis of DNA—measurement of cell kinetics by the bromodeoxyuridine/anti-bromodeoxyuridine method, in Flow Cytometry A Practical Approach, 2nd ed. (Ormerod, M. G., ed), IRL, Oxford, UK, pp 137–156.Google Scholar
  5. 5.
    Latt, S A. (1973) Microfluorometric detection of deoxyribonucleic acid replication in human metaphase chromosomes Proc. Natl Acad Sci USA 70, 3395–3399PubMedCrossRefGoogle Scholar
  6. 6.
    Crissman, H. A. and Steinkamp, J. A. (1987) A new method for rapid and sensitive detection of bromodeoxyuridine in DNA replicating cells. Exp Cell Res. 173, 256–261.PubMedCrossRefGoogle Scholar
  7. 7.
    Rabinovitch, P. S., Kubbies, M., Chen, Y. C, Schindler, D., and Hoehn, H. (1988) BrdUrd-Hoechst flow cytometry A unique tool for quantitative cell cycle analysis. Exp Cell Res 74, 309–318CrossRefGoogle Scholar
  8. 8.
    Ormerod, M. G. and Kubbies, M. (1992) Cell cycle analysis of asynchronous cells by flow cytometry using bromodeoxyuridine label and Hoechst-propidium iodide stain. Cytometry 13, 678–685PubMedCrossRefGoogle Scholar
  9. 9.
    Ormerod, M G., Imne, P. R, Loverock, P, and Ter Haar, G (1991) A flow cytometric study of the effect of heat on kinetics of cell proliferation of Chinese hamster V79 cells Cell Prohf. 25, 41–51CrossRefGoogle Scholar
  10. 10.
    Ormerod, M. G., Orr, R M., and Peacock, J. H. (1994) The role of apoptosis in cell killing by cisplatin, a flow cytometric study. Brit J Cancer 69, 93–100.PubMedCrossRefGoogle Scholar
  11. 11.
    Poot, M and Ormerod, M G. (1994) Analysis of proliferation using the bromodeoxyuridine-Hoechst/ethidium bromide method, in Flow Cytometry: A Practical Approach, 2nd ed. (Ormerod, M. G., ed.), IRL Press, Oxford, UK, pp. 157–167Google Scholar
  12. 12.
    Kubbies, M., Goller, B., and Van Bockstaele, D. R (1992) Improved BrdUrd-Hoechst bivanate cell kinetic analysis by helium-cadmium single laser excitation Cytometry 13, 782–789PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1998

Authors and Affiliations

  1. 1.ReigateUK

Personalised recommendations

Cite protocol

Buy options