Continuous bromodeoxyuridine labeling and bivariate ethidium bromide/Hoechst flow cytometry in cell kinetics.
Most techniques of flow cytometric cell cycle analysis are not capable of distinguishing the number of rounds of DNA synthesis that a cell has undergone since the start of an experiment. Continuous labeling with 5-bromodeoxyuridine (BrdUrd) offers such a potential. We illustrate here that the bivariate analysis of non-BrdUrd-quenched ethidium bromide vs. BrdUrd-quenched Hoechst 33258 fluorescence offers a high degree of resolution that enhances the analytical power of the technique, and that this approach can be applied to the analysis of a broad range of human and murine primary cells and established cell lines.
Poot
M
M
Department of Human Genetics, University of Wuerzburg, Federal Republic of Germany.
Schmitt
H
H
Seyschab
H
H
Koehler
J
J
Chen
U
U
Kaempf
U
U
Kubbies
M
M
Schindler
D
D
Rabinovitch
P S
PS
Hoehn
H
H
D006690:Bisbenzimidazole; D001973:Bromodeoxyuridine; D002453:Cell Cycle; D002478:Cells, Cultured; D004261:DNA Replication; D004996:Ethidium; D005347:Fibroblasts / Q000032:analysis* / Q000166:cytology; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans
D004996:Ethidium; D001973:Bromodeoxyuridine; D006690:Bisbenzimidazole