[Application of simultaneous flow cytometric bromodeoxyuridine (BrdU)/DNA analysis. 1. Basic and technical studies].
The precise in vitro and in vivo BrdU labeling methods and the optimal combination of HCl concentration, treatment time and temperature for DNA denaturation for use in simultaneous flow cytometric BrdU/DNA analysis were investigated. An EPICS C Cell Sorter with computer analyser was used. 1) For precise labeling of exponentially growing MBT-2 cells the cell should be exposed to 10 microM BrdU for 30 min. 2) For optimal labeling of mouse subcutaneous MBT-2 tumor cells 40 micrograms of BrdU per gram should be injected intraperitoneally for 1 hour. 3) The procedure yielding the most quantitative results for DNA denaturation was treatment of the cells with 4N HCl for 20 min at 20 degrees C. Under these conditions, reliable data were always obtained. In the future, this flow cytometric two-color analysis will be applied to clinical fields and further development is expected.
Shimabukuro
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Department of Urology, Yamaguchi University School of Medicine.
D000818:Animals; D001973:Bromodeoxyuridine / Q000008:administration & dosage; D004247:DNA / Q000032:analysis*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D005455:Fluorescent Antibody Technique; D008722:Methods; D051379:Mice; D008809:Mice, Inbred C3H; D009374:Neoplasms, Experimental / Q000473:pathology; D009691:Nucleic Acid Denaturation
D004247:DNA; D001973:Bromodeoxyuridine