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 (for further discussion, see ref. 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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ormerod, M. G. (1994) Analysis of DNA. General methods, in Flow Cytometry: A Practical Approach, 2nd ed. (Ormerod, M. G., ed.), IRL Press, Oxford University Press, Oxford, UK, pp. 118–135.
Dolbeare, F. A., Gratzner, H. G., Pallavicini, M. G., and Gray, J. W. (1983) Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine. Proc. Natl. Acad. Sci. USA 80, 5573–5577.
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.
Wilson, G. D. (1994) Flow Cytometry: A Practical Approach, 2nd ed. (Ormerod, M. G., ed.), IRL Press, Oxford University Press, Oxford, UK.
Latt, S. A. (1973) Microfluorometric detection of deoxyribonucleic acid replication in human metaphase chromosomes. Proc. Natl. Acad. Sci. USA 70, 3395–3399.
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.
Rabinovitch, P. S., Kubbies, M., Chen, Y. C., Schindler, D., and Hoehn, H., (1988) BrdU-Hoechst flow cytometry. A unique tool for quantitative cell cycle analysis. Exp. Cell Res. 74, 309–318.
Ormerod, M. G., and Kubbies, M. (1992) Cell cycle analysis of asynchronous cell populations by flow cytometry using bromodeoxyuridine label and Hoechst-propidium iodide stain. Cytometry 13, 678–685.
Ormerod, M. G., Imrie, 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 Prolif. 25, 41–51.
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.
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 University Press, Oxford, UK, pp. 157–167.
Kubbies, M., Goller, B., and Van Bockstaele, D. R. (1992) Improved BrdUrd-Hoechst bivariate cell kinetic analysis by helium-cadmium single laser excitation. Cytometry 13, 782–789.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1997 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Ormerod, M.G. (1997). Analysis of Cell Proliferation Using the Bromodeoxyuridine/Hoechst-Ethidium Bromide Method. In: Pollard, J.W., Walker, J.M. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology™, vol 75. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-441-0:357
Download citation
DOI: https://doi.org/10.1385/0-89603-441-0:357
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-441-9
Online ISBN: 978-1-59259-561-7
eBook Packages: Springer Protocols